/sci/ - Science & Math

>>9554648
Do it, record results

>>9554648
Very, very, very, very risky. In the best case scenario you actually suceed in modifying your genes and die of an autoimmune reaction. More likely you're gonna waste a bunch of time and money and nothing will happen.

Gene therapy doesn't work except in very limited cases. If it actually worked why don't you think there have been successful medical trials using CRISPR in humans?

>>9554659
>If it actually worked why don't you think there have been successful medical trials using CRISPR in humans?
Because ophthalmologists are the most complacent and incompetent fucks I have ever seen in my life. There's promising research on doing this, but they aren't willing to actually use it.

>>9554668
you're the incompetent fuck wanting to shoot up CRISPR like heroine without doing so much as any testing whatsoever in an animal model.

>>9554681
Of course I would do that first. I only got preliminary results a few hours ago after they forgot to run the sample for five months. They're still "inconclusive" because they're not sure if the mutation is harmful or a natural variant yet.

>>9554648

what is the mutation?

I know some things about the subject, and give you some pointers, and tell you what's possible and what's not.

but I can tell you that you can't "change your dna" the way you're probably imagining it.

>>9554700
I have a previously undescribed mutation in my EYS gene, so they don't know if it's benign or not.

I have really atypical retinal degeneration that started about two years ago. There's uncertainty whether it's genetic or an autoimmune condition, or potentially both. I also tested positive for anti-retinal antibodies and I'm currently on immunosuppressants. Normally EYS mutations correspond to retinitis pugmentosa type 25, but my electroretinogram keeps showing signs of cone-dominated dysfunction despite my perfect color vision. Also it's mostly just in my right eye,

>>9554700
>>9554709
Plus with the retina normally being immune-privileged, combined with my medication, I ought to have some sort of safety net, right?

>implying crispr works like the green goblin serum from spiderman
i'm sorry

>>9554726
They've used it to successfully treat other RP-causing mutations in humans. Just not this one yet.

Are you willing to shoot plasmids directly into your eyeball? The eye is a pretty isolated region from the rest if the body, there's not a lot of exchange of anything except gases

>>9554734
My dad is a nephrologist. The plan would be to talk him into doing it at home while I'm sedated.

>>9554736
Your dad is Not going to help you perform risky home surgery that might cause unintended mutations everywhere in your body, especially when it's not even his field of expertise.

>>9554742
I didn't say it would be an easy sell.

>>9554747
It would be a pretty direct violation of his medical oaths.

>>9554750
It wouldn't be the first time that's happened, but that's besides the point. Can we please keep this thread on topic with respect to applications of CRISPR to retinal diseases?

What disease? I don't see why not.

Might as well. What's the worst that could happen? You fuck up your already fucked up genes? Best case scenario, you cure yourself. Worst case scenario, you kill yourself by basically causing super cancer and letting a bunch of mutated genes run rampant and multiply in your system. Either way, don't be a bitch.

>>9554709
>>9554712

I'm sorry about your condition.

But yes, if it only affects the specialized sensory neurons in your retina then there might be a chance, as long as they don't die.

you definitely need to keep taking those immunosupressants for a while.

you need to figure out whether it's an issue with the histocompatibility complex or something else, and whether the dystrophy is due to cytotoxic processes (e.g. NK cells), in which case you might be able to engineer a virus that can insert a deactivating surface presenting protein producing sequence into the affected cells.

I think it's possible.

You'll need a well founded education in human physiology and pathophysiology, and then bioinformatics. If you're really motivated, you can probably get to where you need to be in 5-6 years.

Genetics thread for retards
>Discussion of methods of genetic engineering and how one would use those methods to make money/a difference
So, i was thinking. AIDS virus changes your dna. Thats pretty fucking cool, make we could rumble up AIDS and make it target not my immune system genes, but other genes, like cancer genes.
Is the cure to cancer AIDS?

>>9554766
>You'll need a well founded education in human physiology and pathophysiology, and then bioinformatics. If you're really motivated, you can probably get to where you need to be in 5-6 years.
Not OP, but how do I obtain this?

>>9554778
>sexually transmitted cure for cancer
I like the way you think.

>>9554766
what the fuck is this shit possible

people can just fix their own genetic mutations like mad scientists now?

>>9554693
Yeah you totally have the funds and capability necessary to conduct a clinical trial of the kind that would give meaningful data. Shut the fuck up and go to a different doctor.

>>9554814
>>9554648
im getting mixed vibes from this thread

is it or is it not possible to cure your own genetic disease by yourself like a mad scientist in your basement?

>>9554814
http://www.the-odin.com/diy-crispr-kit/
$159. Opinion discarded.

>>9554826
with n=1 you can't tell much of anything

>>9554750
>>9554758
Now I see how OP ended up this retarded.

>>9554810
>people can just fix their own genetic mutations like mad scientists now?
it's been possible for a few years

>>9554659
>If it actually worked why don't you think there have been successful medical trials using CRISPR in humans?

Ethics issues.

>>9554818
Possible? Yes, in a I might discover a massive seam of gold under my house way.
>>9554826
Great Job! Now op just needs to somehow buy the education and work experience necessary to conduct a clinical trial of any significance off amazon and hes good to go! Dont forget to add all the lab assistants, and facility cost to cart though.

>>9554766
>>9554814
>>9554863
I already have the bioinformatics part covered. I need to read up on molecular genetics still.

>>9554818
it's possible to attempt it. it is not advisable in the slightest.

>>9554818
The answer is yes, maybe, but you are almost guaranteed to fuck yourself up.

>>9554648
you WILL die

Okay, new plan:

EYS codes for an aqueous protein that is excreted from healthy cells and used to stabilize the surrounding extracellular matrix. Since it's extracellular I should be able to synthesize it in Escheria coli, just like insulin, and then inject it retinally. I've gone 23 years without a functioning copy, so injections every 2-3 years should be curative.

Tell me what's wrong with this plan.

>>9555384
You lack a basic understand if genetics would be the first real hurdle here.

>>9554659
>best case scenario: you die
>more likely: nothing happens

this is why i hang around here

>>9554659
Because leftists despise anything that suggests natural hierarchy like genetics do.

>>9554648
What's the future potential of a CRISPR treatment for crohns? Mode of administration would be easy: either alimentary tract for gut or blood for immune system. The genes behind it are well documented.

So what's stopping it when there are essentially no treatments right now?

>>9554648
>thinks genetics are like LEGOs
you need to stop watching so many hollywood movies.

>>9555384
How much are you going to inject? If it stabilizes ECM, are you going to injectultiple places or just one and hope it diffuses?

>>9554659
H-human TESTING!?!??!?!

Man, i injected myself with nigerian dna, then chopped my dick of so that it would grow back longer, but now im just a retard without a penor.


not advised!

>>9555705
Not to mention you're almost certainly HIV+ now.

>>9555384
>my immune system has a kink and secretes antibodies that form harmful complexes with some aqueous protein
>let's just inject more of it!
I wish being blind didn't preclude you from posting your results here

Don't. Attempt. Your own. Cure.
I'm sure being in /sci/ for too long has inflated your ego and convinced you that the reason your condition (along with most autoimmune disorders) has not been cured yet is because people in the field don't have your smart pretty head, but these things are slow precisely to prevent arrogant and overconfident people like you from harming themselves

Worked for me.

>>9554760
>I don't see why not.

That's cold, man.

>>9554790
>Read a book
>Read a book
>Read a muthafuggin book

>>9555472
More specifically?

>>9555581
Not sure yet. I might be able to get away with just injecting it into the vitreous, or better yet the bloodstream. I have more research to do.

>>9555732
It's not definitively autoimmune at all. I was placed on those drugs prophylactically, and tests for anti-retinal antibodies are notoriously nonspecific for actual disease.

>>9555872
>I might be able to get away with just injecting it into the vitreous, or better yet the bloodstream.
bloodstream won't work, the eye is immune-privileged.

>>9555875
I know, but it's already designed to diffuse through the retina, so I'm not sure if the BRB might permit it to enter.

>>9555758
Which boog?

I had no idea crispr was something you could do in your room

>>9555507
youre are retarded

>>9554810
nope, only works on embryos

>that one time 4chan cured a rare and formerly untreatable disease

Day 2 update:

It looks like EYS is also a CRB1 antagonist, so it's going to be really important to get the dosing right in order to avoid creating even bigger problems like RP12 or LCA. In that case, dosing might work best via a retinal implant like those ones they have for severe cases of uveitis. Of course, without the steroids that give you horrible cataracts after a couple years.

>>9557933
Also starting to look into sources of funding for commercialization btw.

>>9554659
>best case scenario: you die
>more likely: nothing happens

>>9554766
Not OP, thanks for the info.

>>9554810
Halfway between these two:

>>9556737
>>9554852

It's easier in the germline but it is possible to do it somatically, though we'll need better vectors of introduction for that to really be viable

>>9556737
Not remotely true. Embryonic is easiest but then you have stem cell resevoirs of areas of cell turn over such as the gut or bone marrow. Finally you have differentiated cells but these two can be targered, just with a mosaic problem to overcome.

>>9554648

Make sure to reboot your system after upgrading your drivers.

Day 3 update:

Reached out to someone I knew from high school who's a PhD candidate at Stanford now. He has much more experience with this molecular genetics stuff than I do. Ideally I would like his help in developing this protein replacement therapy, but right now I'll settle for his feedback on my approach.

The current outline of my plan is:
- EYS has 6 isoforms, 4 of which are important to the retina, so pre-splice each isoform for transcription
- Insert each isoform into E. coli, which can be used to produce the protein in industrial quantities
- Demonstrate effectiveness in modified zebrafish models (EYS is conserved across the animal kingdom, and the human copy is a drop-in replacement)
- Demonstrate safety in mammals, as well as determine therapeutic dosing
- Begin clinical trials (FDA orphan drug exceptions should speed this along)
- The protein can be injected into the vitreous as an outpatient procedure every 6-18 months
- The vitreous serves as a reservoir for the protein and allows it to slowly diffuse into the retina over time

The main challenges with this approach are:
- EYS is an antagonist of CRB1, so the dosing must be perfected to avoid creating further complications like RP12 or LCA
- Mice have no copy of EYS, so testing is limited to fruit flies, zebrafish, and primates
- EYS is 2.2 Mb long, so I'm not sure if this might present issues with producing transgenic E. coli

>>9560105
Well, I guess best of luck OP.

I hate to say this, but even if you get the ball rolling on this it's gonna be many, many years before you synthesize a cure and then get the FDA to approve it for human use... but at least you can make a contribution for people like you.

>>9560105
>Begin clinical trials (FDA orphan drug exceptions should speed this along)
see you in 10 years

>>9560135
>>9560155
Meh, I'm still willing to bootleg it once I've figured out therapeutic dosing. Plus the FDA has a track record of approving protein replacement therapies very quickly, and this has a minimal risk profile due to being contained in the eye.

Friendly reminder

>>9560460
that guy seems like the quintessential douchebag if i've ever seen one

>>9554659
>and die of an autoimmune reaction
More like get arthritis and be a bit sore

>>9560460
>being a literal soyboy

>>9560526
This meme is getting tired fast.

>>9555507
There is at least one hierarchy we can all agree on, and it goes left-wing > right-wing.

Do you want to die /sci/?? If not get in here:

https://discord.gg/ftSbffu
https://discord.gg/DDpUqJh

I guess now the question is do I buy my own lab equipment, or do I find a way to use someone else's? Thoughts?

Starting out I'm just going to be doing some basic protein synthesis and CRISPR gene knockout in zebrafish embryos.

>>9561559
Make your own thread.

>>9561559
What's in this discord?

>>9561660
Have you considered going to school and making research into this the topic of your dissertation? Could gain you access to equipment, and help depending on your current level of education.

>>9562415
I had considered it, but I'm worried about getting bogged down in irrelevant graduate coursework that would slow down my development of a cure. I probably have about 5-10 years of functioning vision left, so I need to produce this as quickly as possible.

Also I'm not sure how graduate programs would feel about admitting someone with an undergrad degree in an unrelated field. I have a BSc in mechanical engineering and was about to go for an applied math PhD before the vision problems started to hit.

>>9554648
In theory it would but if I was you I would not just mutate one gene I would make a better human

>>9561484
> universities are artificially increasingly left wing
> universities politically indoctrinate students
> people with degrees are therefore left wing
None of that entails a causative link between intelligence and left wing predisposition. In fact a recent major survey found that when tested for political knowledge, people on the right came out much better. Im a doctor and i recall almost all my lectures starting with political asides and that had an impact whereby i had to keep my political beliefs very quiet in order to not run into issues.

>>9562794
https://en.wikipedia.org/wiki/Liberal_bias_in_academia#Implications
>There is little evidence, however, that the political orientation of faculty members affects the political attitudes of their students.[15] A 2008 study by Mack D. Mariani and Gordon J. Hewitt found no evidence that faculty ideology was "associated with changes in students' ideological orientation" and concluded that students at more liberal schools "were not statistically more likely to move to the left" than students at other institutions.[12] Similarly, Staneley Rothman, April Kelly-Woessner, and Mathew Wossner found in 2010 that students' "aggregate attitudes do not appear to vary much between their first and final years," and wrote that this "raises some questions about charges that campuses politically indoctrinate students."[16]
But I suppose Wikipedia is run by liberals too, so you can't trust that.

Day 4 update:

Pirated/ordered some books. Talked with my contact at Stanford. Starting to work out some of the implementation details and build out a support network of people who have more experience doing this than me.

>>9562794
>>9562812
Take it somewhere else.

>>9554668
there's promising research but they arent willing to SEE IT

HAHA LOL HEARTY KEKS

>>9554736
basically this:
>>9554742

your dad does not want to puncture your eyeball, then see you get super sick from a failed mutation, and then morn your death as he get gang raped in a prison cell because of the huge legal issues hell get into

>>9554808
actually, if anyone ever invented some kind of super benign healing virus, this would be the best way to make it spread

>>9554778
the virus is HIV

>>9562883
Please keep us updated. Maybe start a blog or something and link it here.

>>9562812
No but a university research study on the bad effects of left wing university professors is clearly going to be useless out of the gate.

>>9565307
Clearly.

>>9554648
kek

>>9560105
>he is actually going through with this

>>9554648
this should be good.

Day 4 update:

Spent most of the day trying to reverse engineer wtf this little guy actually does. Without having run actually experiments yet, I'm trying to make an educated guess about its function by looking at similar proteins and trying to reconstruct how it might have evolved, and then corroborating that with some of my symptoms (e.g. lots of flashing lights with vigorous exercise, flashes go away when holding breath/lower blood pH).

My current best guess is that it uses Ca2+ ions to facilitate F-actin remodelling, but I think there's more to it than that. I have a feeling that it also does some sort of calcium scavenging, because muscle contraction tends to release free calcium ions, and holding your breath ought to reabsorb them (plus I started drinking an assload of milk around the time this started getting bad). It's also descended from epidermal growth factor (EGF), which probably means it has some additional role in cell signalling or gene expression. I think the next few days will be pinning down what EYS does a bit better.

Day 5 update:

Spent most of the day trying to reverse engineer wtf this little guy actually does. Without having run actually experiments yet, I'm trying to make an educated guess about its function by looking at similar proteins and trying to reconstruct how it might have evolved, and then corroborating that with some of my symptoms (e.g. lots of flashing lights with vigorous exercise, flashes go away when holding breath/lower blood pH).

My current best guess is that it uses Ca2+ ions to facilitate F-actin remodelling, but I think there's more to it than that. I have a feeling that it also does some sort of calcium scavenging, because muscle contraction tends to release free calcium ions, and holding your breath ought to reabsorb them (plus I started drinking an assload of milk around the time this started getting bad). It's also descended from epidermal growth factor (EGF), which probably means it has some additional role in cell signalling or gene expression. I think the next few days will be pinning down what EYS does a bit better.

Will CRISPR beckon the dawn of anime catgirls?

>>9565519
This in all honesty is just a reflection of mass immigration into white countries. It's not showing where evolution is taking us so much as it's showing what is happening/will happen to white genetics as it is polluted with non-white genes.

>>9560105
>Mice have no copy of EYS, so testing is limited to fruit flies, zebrafish, and primates
>primates
may I suggest to abduct random hobos to experiment with?

>>9564575
I'm considering it, but I probably wouldn't want to link my identity to 4chan. There's way too many underage edgelords that will go out of their way hurt you.

Also another thing to consider is that if I'm going to have to buy lab equipment, then I'm going to need funding, and that might mean investors. If I create my own prior art by hosting a blog then I might not be able to secure a patent and they won't to give me money. It fucking sucks that the only way ((they)) will let me make this is by charging insurance $100k per dose because so few people have it.

>>9565553
Only if I can get to them as embryos and do gene knockout.

>>9565552
the fact that less intelligent people tend to reproduce more should tell you something too.
Suburban & rural retards spout 100 kids each while educated city folks rarely have more than 1, if at all.
It's the consequence of modernity and hedonism.

ITT: OP is just smart enough to think about fixing his genetic disease, but just dumb enough to forget that DNA\RNA genes operate in a cyclically interdependent manner, such that it is currently impossible to predict the effect of manipulation of one or multiple genes in a complex animal. The DNA\RNA system operates on rules which chain and combine the effects of codes in a manner that has and will tend to defy analysis. This is why there is no truly abstract field for animal husbandry, as it's been based on experimentation which cannot be properly quantified.

tl;dr - Nobody knows what happens when you change someone's genes, because it's fucking ridiculous how DNA works. You can pretend that you could fix yours or someone else's genes, but there's absolutely no way to predict what genes will interact with the genes you alter, but for experimentation--with all the limitations associated.

Do what you want to do, anon, but do please post your strange eczema and buboles before you go blind anyway.

>>9565584
Maybe you should read the thread. He's not doing gene therapy.

>>9565589
Yeah, "protein replacement therapy"

Care to explain the difference, given interactions?

>>9565596
You're too dumb to post here. Go away.

>>9565597
>posting on a board with moon landing denial threads

>>9554659
You gonna replace ALL the DNA in ALL your bodies cells?

How?

>>9555384
Dont listen to the haters OP. They were saying the same shit about homemade 3D printed tooth aligner anon on /diy/ a few years back. Who’s laughing now? Invisalignanon is, because he has perfect teeth. Just make sure you do your research and dont just inject useless garbage into your eyes.

>>9565565
What is with the shilling for living in the city lately? Why would i pay more money for a smaller house on less land, with flouridated water, polluted air, and niggers all around it? Thats what stupid people do.

>>9560499
>change every cell mhc genes
>get arthritis and be a bit sore

do it OP, become the madman scientist you always wanted to be.

>>9565726
reminder that 4chan is the most influential place on the internet. we elected the president

>>9554808
Would it be possible to make a sexually transmitted form of the HPV Vaccine?

>>9567284
Why did you say this in response to my post? I cant tell if are are supporting me or telling me to stop.

you will fuck up and your dick and balls will riot off then your legs and then arms.

>>9554648
your best bet is steam cell therapy by intravenous. panama mexico has american docs doing it. costs about 30g

Day 6 update:

I think I have a much better understanding of what EYS does now. It's a proteoglycan that selectively binds to prominin receptors that are expressed exclusively on the photoreceptor outer segments (the part of the cell that holds the pigment granules and does phototransduction). EYS forms a sort of cell wall around the rod/cone outer segment that protects it from osmotic pressure during heavy environmental stress. This probably explains the bit about my sensitivity to blood calcium levels, where I'm physically deforming my photoreceptors due to osmotic forces, damaging them in the process.

I'm encouraged by the fact that EYS likes to bind to prominin, which is only expressed specifically where I need it to go. However, it's a little discouraging that it's a proteoglycan. This might mean that it ends up crosslinking to itself in solution, as well as any other random actin fibers it comes across, before it can diffuse into the photoreceptor outer segments. I might be able to inject it into the subretinal space so that it's right by where it needs to go, but that seems risky. Also I don't want to clog up my Bruch's membrane with this stuff and cut off oxygen to my retina. I need to do more reading.

Also, I'm going to email some NIH researchers tomorrow to get their opinion. I'm trying to have a detailed proposal written up by the end of the week that I can show to donors/investors, letting me start to procure lab equipment.

>>9567910
God speed OP. The haters are just jealous that you are using science to help yourself instead of the mental masturbation they have done. Keep doing your research and keep us updated. This thread could become legend.

>>9567910
Good luck OP.

>>9567910
Very interesting about the calcium sensitivity. Bumping your thread so it doesn't get archived before you come back. Should it ever get archived, I (and maybe others) would appreciate you making another thread (with some sort of identification for us to easily find it) or keep us updated on the /sci/ archive at warosu.org/sci. Here's your thread in particular: >>/sci/thread/S9554648

Also, I have a question regarding the eye, since you seem knowledgeable. I have what's known as "eye floaters" and they're quite annoying. These come with a lot of other side effects such as flashing lights (ophthalmologist confirmed that my retina is OK), object outlines appearing more readily when staring at an object, and a black flickering blob appearing in fmy ield of view when turning my head and eyes fully sideways. Could you tell me anything about this?

>>9567910
Good luck OP, keep us posted. While getting an approval from FDA seems impossible before CRISPR goes live in medical care, it's fascinating and you could find professionals interested

omg
just do it (if life doesn't matter anymore)
the results could be amazing

>>9568810
>These come with a lot of other side effects such as flashing lights (ophthalmologist confirmed that my retina is OK), object outlines appearing more readily when staring at an object, and a black flickering blob appearing in fmy ield of view when turning my head and eyes fully sideways.
I'm a mechanical engineer and not an MD, but yeah that definitely does sound familiar. I'm a bit reluctant to say that everyone who see weird stuff has the same rare disease I do, but I guess follow up with them and ask if they see any retinal arteriole narrowing or bone spicules. Those are pretty indicative of retinal degeneration.

>>9568250
>>9568276
>>9568852
Thanks anons, I appreciate it.

Day 7 update:

Spent some time with the gf and didn't get as much done as I had hoped.

I don't think premature crosslinking should be as much of an issue anymore. EYS forms a nonreactive dimer with itself until it comes into contact with prominin surface receptors. So really now the only potential issue is that prominin may also be expressed to somewhat on the neighboring Mueller glial cells. It's not clear to what extent EYS will bind to the Mueller glia, or whether it depositing a carbohydrate cell wall on them would be an issue. I'll have to experiment in the zebrafish about doing vitreous versus subretinal injections. If EYS clings too much to the Mueller cells then I'll just have to bypass them by administering subretinally.

I'm starting to look into the details of how to produce the protein now. Also I still have to send that email.

Day 7 update part 2:

I forgot to mention that the time spent with my girlfriend mostly involved me holding her hand as she got fucked, hard, by her bull. Yes I'm a cuck. Call me whatever you want, a blind cuck even but you're just immature. We love each other and soon with genetic editing i will remove my ability to hear your judgements.

>>9570289

epic

>>9570289
Actual OP here, that's not remotely me and thanks for trying to derail my important thread. Probably one of the only important things /sci/ will ever accomplish and you try and ruin it with your cuckposting. Why don't you stop caring about what other people do with their love lives? Being so obsessed makes you the real cuck, not the guy having fun with his girlfriend.

>>9570304
Ignore the trolls OP. Make a trip number, that way you wont even need to clarify what is you and what isnt. Normally trips are gay but this is the exact situation where they aren’t. Keep up the research, sounds like you have learned a lot already, but there is no such thing as too much knowledge when it comes to editing your own genes. Good luck you maniac.

>>9570304
But you're not me either.

>>9570545
Okay this is my trip now.

OP is this from birth or did you only just start getting symptoms? If it's recently and actually has an autoimmune pathogenesis, you can always try helminth therapy, probiotics, or mesenchymal stem cells.

Day 8 update:

Whelp, found out what those other isoforms do. Apparently EYS is also expressed in the spinal chord and dorsal root ganglia to prevent osmotic shock there too. That explains the nerve problems I've been getting, which I had thought were from my medication. As of today I'm drastically increasing my water consumption (1-2 glasses an hour), as well as restricting my salt and refined sugar intake. Hopefully that will help me maintain my osmotic pressure a bit better and buy me some more time. It's too early to say, but it does seem like making that change has reduced the flashing lights somewhat already.

Also reading more about all of my options for producing the protein. I haven't settled on anything yet, but there's a good chance that I'll end up using chinese hamster ovary cell cultures. CHO should be able to properly glycosylate EYS after transcription, and they should also make regulatory approval easier.

With introns, EYS is 2.2 Mb long, but without it's only about 11 kb, so I'm going to make a cDNA copy of mRNA that's expressing it from human retinal cells. Then it will actually be small enough to insert into my cell culture.

>>9571889
I had a few brief, isolated flare-ups over the years (5th grade, freshman and senior years of high school) but they always resolved pretty quickly and I didn't think much of them. Then two years ago something happened and it just started going crazy and never let up.

So until they run those confirmatory genetic tests on my parents, I'm still not 100% certain one way or the other. But in the mean time I might as well get prepared for it being genetic.

>>9554648
chances of success are very slim.

>>9572445
From the looks of it OP is taking this seriously so he still might get interesting results even without curing the condition.

>>9572422
Post pics when you start setting up.

>>9555484
Jej

>>9561484
Fucking extremists. You take away my oxygen

>>9555525
I want to know this. My father has it

>>9572575
Sure, but it'll probably be a few weeks to a month before I get lab equipment. I have to obtain funding first.

>>9554659
What makes you think the Chinese already haven't created a large subpopulation of supermen?

>>9555384
>Tell me what's wrong with this plan.
How long you got?

>>9573102
Best of luck, but keep us updated through regular threads please. Would be great to be able to follow your progress.

>>9573114
Probably about five years. Maybe more if I can figure out a way to manage the progression better.

>>9573117
Thanks anon, will do.

Day 9 update:

Still reading about ways of producing the protein. The current plan boils down to
>isolate mRNA from retinal cells
>use reverse transcriptase to convert to CDNA
>amplify CDNA using PCR
>transfect CHO cells using cDNA and electroporation
>make the changes permanent using CRISPR
>screen the resulting cell colonies
>verify that their copy of EYS made the transfer safely via sequencing
>begin production and purification
>???
>profit!

Starting to price out equipment too. I think I can do the first zebrafish trial for $15-$25k if I borrow some things.

Also I emailed those researchers.

>>9574832
Where are you planning to get the funding needed?

>>9574837
A few possibilities:
> NIH research grants
> crowdfunding
> investors

I had a roommate in college whose dad did early stage investing in startup companies. At the time he had offered to give me money for a different project, but I never took him up on his offer. I might contact him again.

>>9575459
If I were you, I'd try that. Probably the most surefire way. Sadly I can't offer any advice on the scientific side as biology isn't my strong suite. Still interested to see how this turns out. Good luck and have fun.

>>9554709
Put an eye patch on and invest your money in the balkans.

Day 10 update:

It's becoming clear that producing a viable cDNA for EYS is going to be the main challenge here. I'm currently exploring the best way of doing this. If I can use artificial DNA synthesis without introducing too many transcription errors, then that would make my job waayy easier

>>9577109
Forgot trip.

>>9575685
Thanks anon! I want to have all the details worked out beforehand, as well as a required bill of materials. Then I can just show them exactly what I'm going to do and how it will work.

>>9576380
What did he mean by this?

>>9577110
Did those researchers that you emailed answer?

>>9578017
Not yet. I emailed them after they would have already left the office Friday evening. Hopefully tomorrow or Tuesday though.

>>9579198
Do you have any background in science? I can guarantee you the NIH is going to think you're a crackpot if you don't.

>>9567296
That's a kind of Russian Roulette I won't be ready to play. Especially as someone carrying HPV 51 and 55, probably since birth.

>>9579198
You are an absolute raving madman. I like you and hope you succeed.
And do remember that proving a hypothesis wrong is success too.
Meanwhile I'll just keep scrolling through papers looking for research to base my artificial eye startup on.

Day 11 update:

Figured out how I'm going to genetically isolate EYS. First, I'm going to make a cDNA library of human retinal cells. That means popping the cells with an enzyme so that they spill their guts and messenger RNA into solution, then using reverse transcriptase to convert the mRNA into cDNA, then amplifying the cDNA with polymerase chain reaction, before finally inserting all the random cDNA genes into a hamster ovary cell culture using electroporation. Once I've made the library, I'll integrate all of the plasmids into the CHO germline using CRISPR/Cpf1. I only want the CHO cells that integrated EYS cDNA, so then I'm going do a second round of targeted CRISPR/Cpf1 editing to confer resistance against methotrexate, where the guide RNA will be targeting EYS. Only the subset of cells that are producing EYS will have resistance, so then I just dump methotrexate into solution to kill off all the others. Plus, this lets me skip a step since higher methotrexate resistance will be correlated more copies of EYS and thus higher protein productivity.

Also pic related came in the mail today. It's pretty basic, but I'm only using it to get familiar with the terminology so that I can look up all the advanced details on my own.

>>9579220
Yeah, that's sort of what I'm expecting to happen, though I have to at least give it a shot. Like I mentioned before, I'm a mechanical engineer and I had an offer to do an applied math PhD at Columbia. Fixing my vision is my main priority right now though, so grad school can wait.

>>9579220
Thanks anon! At the very least I hope this spurs more RP25 research since it's currently very understudied.

Also how far is your development and how are you interfacing with the optic nerve? I thought the main bottleneck was getting data to the brain, not the image sensor itself.

>>9579837
I've been considering it for a while, but my day job in developing market 4Kp60 cameras kinda takes priority now - but also gives me some hope. Recent devices do seem to get to the level of miniaturization that would allow to place appropriately-sized wireless camera in the eyesocket as is. Long-term power supply is one problem, SoC CPUs radiating a lot of heat is another, but we might get there in terms of the size of camera module itself without going beyond market capabilities in 2-3 years, really. That brings me to the interface part.

Good grief... The whole optic nerve stimulation setup is finicky as hell, however you look at it. The nerve has throughput like nothing else in your body for sure (my teachers told me that you can get 8 Gb/s in there, but as far as I can see it ~200 Mb/s is our limit here), but it is a bitch to deal with. Implanting penetrating electrode arrays, supplementing with surface electrodes - all of that, as seems from most papers I've seen, works, but doesn't get anywhere near actual carrying capacity of the nerve - because of high-latency ion-based signal propagation in ganglia at the very least. So, we can either:

>>9579957
(cont.)
>Do what these Optogenetics madmen do and genemod the patient with ChR microbial opsins to have light-sensitive ganglia and fuck his shit up with a laser-powered optic fiber array, hoping to get the frequencies just right for the visual cortex to process
Yeah... No. I don't think I have to explain it to you. Hint: CRISPR won't help. You gotta grow your patient with that. Also, lasers are bitches to work with in such miniature setups.

>Limit ourselves to (at the very best, as it seems at this point) 1024x1024 pixel resolution with worse latency than normal eye
What's the point even.

>Stimulate the retina instead of the nerve
That's only half a solution. Also, Chinese are doing it and they're much smarter than I am (got my Biomedical Engineering Master's degree in China, btw)

>Screw the slow-ass neurons, let's bypass it and go straight to visual cortex
Now we're talking. Stick a camera in the socket, implant a bigass platinum microarray into the back of the patient's head with a wire running out of his neck, give him a nice pendant or collar as the modulation device, connect it back to the camera through your wireless network of choice. However... Yeah, you probably guessed it. Who the hell sends an HD signal straight into the brain and hopes the poor sod will discern anything from it? Well, these guys most certainly do: http://www.jneurosci.org/content/jneuro/36/8/2406.full.pdf (not HD, and not even visual spectrum, but the idea holds water for now)
There's a limit to that, of course - and a lot to discuss. But the biggest moot point is the whole ADULT NEURAL PLASTICITY situation. Can a grown brain to a completely new source of information going right into it's cortex? Probably. Maybe.

I don't see a definitive answer yet. And that saddens me.

good thread bumping

>>9555732
On a fun note, pretty fucking worried I've managed to acquire A in that pic.

>>9579957
>>9579969
What do you think about this approach (it keeps saying it's spam)? They solve the heat dissipation, power consumption, resolution, and interfacing issues by using a passive photovoltaic implant that mimics form and function of the photoreceptors. Currently it can only see in ultraviolet, but they talk about how they can dope different nanorods with different concentrations of nanoparticles to sensitize them to different wavelengths, enabling color vision.

https://www.nature.com/articles/s41467 dash 018 dash 03212 dash 0

Testing better trip.

Testing better trip.

>>9580659
Yeah, that's the one I actually inferred in "stimulate the retina" group. I deem it 'half a solution' because it requires a lot while achieves a little:
>Vision restored - and possibly with decent resolution as well
But
>Requires the eye itself to be intact (ruling out using this tech as a solution to traumatic eye injury, glaucoma, etc.)
>Requires the optic nerve to be intact (ruling out degenerative nerve conditions, optic nerve trauma, etc.)
>Currently only sensitive to blue and UV, that is - higher energy light & threshold of light response at 25 lux
Say goodbye to candlelight and your bedside lamp, that's hella weak. To get their concept on level with our normal vision they'll probably have to find a completely different set of materials for them nanowires - of which I haven't heard yet.
They have a nice and tidy technology that could help with a very narrow range of conditions - no external energy supply, no heavyweight implanted devices, but also requiring a very delicate and rather worrying kind of surgical procedure.
Concept I hope to get going is much more bulky, but also significantly more versatile.

Day 12 update:

Apparently you can buy EYS in microgram quantities, but I'm not sure if that's enough to conduct preliminary studies in zebrafish. I need to see if I can get it in greater amounts.

Also, I'm trying to work out the protein purification process a bit better. It looks like the best way to do it is with affinity chromatography, but I'm not sure which variant of this process to use yet. In either case, you bond some type of binding agent to the surface of these porous gel beads so that when you pour your solution through, the protein gets left behind on the beads and everything else filters through. Then to get the protein you wash it off with a special elution agent that causes it to let go.

The easiest way would be to bond some polyclonal EYS antibodies I found to the gel, but one of the papers I had read earlier said that they weren't very good. The alternative would be to introduce a tag to my recombinant EYS protein (maybe a polyhistone tag, maybe something else). It would be a small dangly bit that would get appended onto the end of it during the second CRISPR edit, so that it could bind to a nickel-based affinity chromatography resin. The issue with doing it this other way is that I then have to go back and cleave off the tag with another enzyme that I have to also filter out. Purification is going to be really important so I need to study this some more.

In other news, there may be programs that I can apply to get the FDA to fund human trials themselves, as well as lots of other perks like expedited review and 7 year orphan drug exclusivity. I need to finish drafting this proposal document by the 27th when I have an appointment with the local university's chair of ophthalmology.

>>9582333
>I have an appointment with the local university's chair of ophthalmology

So I take it that those emails have been answered.

>>9581905
I hope you're successful anon! Though I will admit that I got pretty excited when I read that other paper since it comes very close to a practical artificial retina.

>>9582348
Not exactly. My case is so weird and everyone else has messed up so badly that they dumped me on him for liability reasons. Still, he does a lot of research and has labs, so I'm going to run this by him.

>>9582333
*polyhistidine, not polyhistone

>>9582359
How did they mess up? Gave you the wrong treatment?

>>9561439
no it's not

Bumping for interest.

>>9582974
Pretty much everything that could have gone wrong has gone wrong. Multiple incorrect diagnoses, as well as botched diagnostic tests where I was at risk of personal injury. I won't go into all the different details, but one of the worst ones was the fluorescein angiogram.

As I was filling out the release of liability paperwork for possible severe allergic reactions, I warned them that I have a lot of bad allergies to vegetable products like the one the dye is based on. They just kind of laughed and went along with it anyway until I started to get a reaction. Then they had to give me a bunch of intravenous diphenhydramine.

>>9585483
>They just kind of laughed and went along with it
Dafuq. Do they just assume all of their patients are paranoid hypochondriac loonies or something? At the same time, you could've easily demonstrated that you were allergic to it by dripping some on your skin or something.

Day 13 update:

Slight change of plans.

It looks like there are several suppliers of not only purified EYS protein, but also pre-made EYS cDNA, mammalian expression vectors, and even lentiviral vectors. The best part is that they're all really cheap. I can get 3mg of pure protein for $500, which is enough to start small-scale experiments in zebrafish. So with this in mind, I'm switching things up a bit.

My plan is now:
> crowdfund $5k for reagents and other basic equipment
> demonstrate a very minimal proof of concept in zebrafish
> get venture capital funding based on evidence of effectiveness
> produce the final therapeutic protein that I've been describing so far
> do a phase 1 preclincal trial in zebrafish with better experiment methodology
> do a phase 2 preclinical trial in either rabbits or cats
> submit FDA Investigational New Drug application

This means the timeline to my first experiment is now expedited, since I can start small-scale tests much more cheaply than I had originally thought. I'm reformatting my proposal as a crowdfunding application (I won't say where because I don't want to be doxxed), and I hope to start putting hardware together in the next two weeks. I want to determine whether this protein replacement therapy approach is feasible or not as soon as possible, before I sink the next three years of my life into developing it.

I'm going to throw together an electroretinogram machine from a salvaged camera flash bulb and some microcontrollers left over from college. It wouldn't pass peer review since it's going to be really crude, and the A/D converters on the microcontrollers aren't the best, but I just need to demonstrate a difference in waveform amplitudes between the treated fish and controls. Then I can show that to investors to get funding for a better setup.

Day 13 update:

Slight change of plans.

It looks like there are several suppliers of not only purified EYS protein, but also pre-made EYS cDNA, mammalian expression vectors, and even lentiviral vectors. The best part is that they're all really cheap. I can get 3mg of pure protein for $500, which is enough to start small-scale experiments in zebrafish. So with this in mind, I'm switching things up a bit.

My plan is now:
> crowdfund $5k for reagents and other basic equipment
> demonstrate a very minimal proof of concept in zebrafish
> get venture capital funding based on evidence of effectiveness
> produce the final therapeutic protein that I've been describing so far
> do a phase 1 preclincal trial in zebrafish with better experiment methodology
> do a phase 2 preclinical trial in either rabbits or cats
> submit FDA Investigational New Drug application

This means the timeline to my first experiment is now expedited, since I can start small-scale tests much more cheaply than I had originally thought. I'm reformatting my proposal as a crowdfunding application (I won't say where because I don't want to be doxxed), and I hope to start putting hardware together in the next two weeks. I want to determine whether this protein replacement therapy approach is feasible or not as soon as possible, before I sink the next three years of my life into developing it.

I'm going to throw together an electroretinogram machine from a salvaged camera flash bulb and some microcontrollers left over from college. It wouldn't pass peer review since it's going to be really crude, and the A/D converters on the microcontrollers aren't the best, but I just need to demonstrate a difference in waveform amplitudes between the treated fish and controls. Then I can show that to investors to get funding for a better setup.

>>9586213
Yeah, it's really messed up. That fluorescein incident doesn't even scratch the surface of all the bullshit I've had to put up with. My theory is that most of their patients are lot more disabled than I am, so they've developed some kind of superiority complex or something. The waiting room is always filled with geriatric patients, and never anyone else my age.

I couldn't have tested the dye beforehand though, since it was administered by an injection.

>>9587350
Are you delusional?

>>9587350
What background do you have? Do you even know what u are doing?

>>9587350
Have you sequenced your EYS gene to see what mutation/variant type you have?

>>9554648
Very unlikely that CRISPR and Cas9 enter your affected cells unaided. Other than that, they are bacterial components and it has been shown that our body has antibodies against Cas9 already. This means you will probably incite an immune response to the foreign protein and genetic material you just shot into your body, which will make it all useless. Then even inside the cells they need to find their way to the nucleus and edit the correct gene. This is all that CRISPR does, guide Cas9 to a gene. Cas9 can then cut the DNA, which leads the natural DNA repair mechanisms of the body to either stick the strands together (deletion of the gene altogether) or to insert some random part in there. Gene swapping is still in its infancy, all we can reliably do is deletion.

tldr: Good luck anon, waste all of your money on this to finance actual research, please

>>9587350
This is a really optimistic plan anon, why do you think researchers study for so long? 5k for equipment and reagents is realistic enough, however how will you ensure that you can work in sterile conditions? Not doing so would make anything too dangerous?

If you want, I am a pharmacology major with focus on drug delivery. I can look through your proposal and point out flaws, oversights and such. I don't believe your plan will work (it's hard enough to net funding as an established researcher right now), but it's the least I can do. I don't really understand how injecting EYS directly onto the retina instantly improves your extracellular matrix there (haven't read the things you did), but have you thought about immunogenicity? Also intravitreal injections have a problem with macrophages simply clearing any protein you inject, how will you tackle that?

My advice: Don't waste time/resources doing this yourself. If it was so simple as to supplement the defective protein, someone would have done so already, do more literature research (clinicaltrials.gov, pubmed, etc) and try to find research groups focussed on retinal disease. I worked with some people in Finland who examined delivery of nanoparticles to the retina for treatment of Macular Degeneration, so there are researchers out there.

>>9588851
Trust me man, I read a lot of books and many articles, I know what I'm talking about. I absolutely do not need your help, because you appear to be arrogant.

>>9589033
No trip = probably not OP

>>9587350
>I won't say where because I don't want to be doxxed

I understand your concern, but could you at least drop a hint as to where your campaign could be found. Would be interested in donating (if I can get money to spare).

>>9555732
This man, only on /sci/ would a freshman think they are smarter than the doctor

>>9565584
we have all this fancy maths surely we can predict how DNA works

I want OP to go along with this. Just to see another retard killing themselves, thinking they're better than doctors and scientists who still have reservations about using a risky and experimental technique to modify certain genes in a grown human.

>>9587950
No, just extremely motivated.

>>9587955
I'm a mechanical engineer. I do a lot of integrated hardware/electronics/software stuff for robotics. I was going to get an applied math PhD before the vision problems hit.

>>9588239
All I know is that I have two different missense mutations, one in each copy. No idea what they actually are or whether or not they're disease causing yet. They're sequencing samples from my parents now to confirm whether that's causing my vision issues.

>>9588718
I completely agree. That's why I'm doing protein replacement instead.

>>9588851
All I'm attempting for this initial test is CRISPR gene knockout in zebrafish embryos, followed by injecting off the shelf EYS into their vitreal chamber, and then seeing if there is any difference in the visual decline of treated vs. untreated fish, based on the electrical signals recorded by the ERG. The trickiest part is going to be the gene knockout, but even that isn't so bad since they deposit eggs that can be easily manipulated in vitro.

I would really appreciate any constructive criticism though!

EYS is localized by prominin-1 surface proteins and forms a cell wall around certain types of nervous tissue, reinforcing it against osmotic shock. By replacing it, at a minimum I hope to prevent the progression of further damage, but I may also be able to rescue the damaged photoreceptor outer segments by relocalizing their organelles that get scrambled from repeated osmotic insults.

Theoretically speaking, the eyes are supposed to be immune-privileged, so immunogenicity shouldn't be issue. I'm beginning to lean towards using an HEK293 expression vector for better glycosylation over CHO though, and that should help with immunogencity as well. Macrophages aren't supposed to be present in the vitreous, but would you be able to link to some papers where I could read more about that phenomenon?

RP25 is really under-researched though. It was unknown until 2008 which gene was responsible for it, and nobody had a good idea of what EYS actually did until about a year ago. So the time is now to start working on a treatment, and if nobody else is willing to do it then I'll have to take the initiative. I know that I can't bring this to market by myself, but all I need right now is the simplest possible demonstration that a protein replacement therapy is feasible. Then I can attract outside funding and assistance.

>>9589048
It's not up yet, but I'm sure if you search hard enough for it then you should be able to find it.

>>9590325
Thanks, please tell us when the campaign is up tho.

>>9554648
It's not fucking CREAM. It's a screwdriver waiting for the right screw.

>>9577110
>What did he mean by this?
he's encouraging you to become a supervillain

>>9589060
he doesn't think he's smarter. he's motivated and willing to experiment. what do you care if he fails? do you have a vested interest in medical exclusivity?

>>9590325
https://www.sciencedirect.com/science/article/pii/S1350946216300635?via%3Dihub

Like I said I haven't researched this as much as you, so godspeed Eye-anon. Just don't underestimate all facets of getting the protein to where it needs to be. Shit's intricate man. Also apperantly Rabbit eyes are really close to human, so after zebrafish consider using those. The paper I linked was conducted by a group I worked with, they used rabbit exclusively

Bump

Day 14 update:

Out with the gf again: didn't get as much done.

Still working on drafting the crowdfunding submission. I'm trying to make some easy to understand graphics for random potential donors without much background knowledge. Also reading some more about standard procedures for handling zebrafish in experiments, since someone will inevitably bring up animal cruelty.

Other than that, I mainly just need to finish working out all the finer logistical details for this first experiment. There's a lot of them, so that'll probably keep me occupied for the next couple weeks.

>>9590343
Probably within the next few days. We'll see though.

>>9590668
Thank you for this! This paper looks like a really good resource. It's going to take me a little while to digest all of it, but I think it's going to help a lot.

My only issue with rabbits is that mice are one of the few animal species that don't have a copy of EYS, so I wasn't sure if rabbits were too closely related to them. Rabbits are definitely easier to work with, but I don't know yet if I'll be able to use them as models.

>>9592118
What are the laws on animal trials in your country? Will there be legal barriers?

For something like this to work, you would have to this for every cell in your body right?

>>9554652
Good idea no ethical issue if he does it to himself

>>9590444
I can see this whole story becoming a movie.

>>9592095
I should clarify about my girlfriend, we're spending a lot of time together because we've been trying to resolve some issies lately. When she heard about my health condition it was very hard on her and she made a mistake with an ex of hers. That on top of my health news hit me hard, i said some regretable things and she didn't talk to me for a week. That's when i started this thread. Since then she got back in contact and we've been working it out. She's really on board and hopeful that it will solve everything but she's a bit worried about the cost of it all, i hadn't thought about that at all, so im going to reach out to reddit too in order to get as much help as possible before moving to the lab stages.

>>9592095
I may be a cheapskate but i'd fund this.

>this dude is gonna become a real life doc oc
please exclude me from your kill list oh ye mighty

>>9592830
>made a mistake with an ex of hers
>she didn't talk to me for a week
op i hope you aren't being too naive with her

>>9592208
I'm in the US, but I've just adapted my sleep schedule to avoid sunlight. Regulations on fish are pretty relaxed, so it's mainly a public outreach thing.

>>9592272
Nope, just the retinas. I may want to try treating my spinal cord with it years down the road, but my neuropathy is a lot more mild than my vision loss.

>>9593109
The only people on my list are edgelords who try to impersonate me >>9592830 >>9593111

>>9554659
>Gene therapy doesn't work except in very limited cases.
the fuck is immunotherapy then?

>>9554766
Recomended mooc courses? Books?

>>9593321
So you sleep mostly during the day? How do you manage your worklife and appointments to those university professors, then?

>yfw some mech eng /sci/tard makes groundbreaking advancements in genetic medicine

>>9593722
There are plenty of tards like him. Look up "biohackers". One thing they have in common is very shallow understanding of biochem, little to no understanding of organic chem and very naive image of the process of going from an idea to a viable therapy.
He's in for a series of big letdowns. But it's always good to see people learn, so godspeed to him.

>>9594031
he might get lucky. a lot of discoveries happened either en jest or accident.

>>9554648
Crispr is not a new thing. I heard a rumor about several perpetual trial that have been been going on. It focuses around white supremacists in America and foster care parents or legal guardianship with legally unknown parents for business safety purposes. Basically they were being used for human experimentation until parents found out. Kids get features that are unknown in their nuclear family. So crispr has been used unbeknownst....they white peoples excuse was the over use of the word genetic diversity.

>>9594756
They keep getting taken out. Apparently the first cases happened around people of the Dominican Republic as most Haitians don’t exist no more and whatever is there is straight from Africa whom have bought identities. Many are undercover agents for African entities that hate people with straight hair.

>>9594773
Other cases involve Chinese. Not just white supremacists. The problem with making it public is mass hysteria. Chinese have one child policy therefore their arrests are hush otherwise they’d scream out victim. Black people have their civil rights movement. Whites are usually rich doing it for companies.

>>9561484
These "studies" are cooked up by asking people questions about topics and deriving your political leanings from those answers. They don't ask you who you voted for they ask you if you support gay marriage and you stance on guns and so on and so forth. Blacks and hispanics tend to have lower IQs and are hyper conservative in many ways, for example both groups tend to really hate gays, but vote left wing for free shit. This allows the commies who do these studies to flood the right wing side with 80IQ knuckle draggers who are de facto left wing, but have some right wing leanings. Also

>leftists worship IQ when it finds that they are smarties and right wing hitlers are dumb poo poo heads
>leftists suddenly start stuttering about how IQ is deeply flawed when blacks are found to be hyper violent literal retards

>>9594785
I am not gonna spoil your fancy conspiracy theory as a whole, but I wanna point out that 'one child policy' has not been a thing for years now; and was never a thing in rural China.

>>9594756
It sounds like you are shedding IQ points by the second.

Day 15 update:

Reading a ton about retinal pharmokinetics. The fact that EYS is negatively charged should be really helpful for having it diffuse throughout the vitreous and retina. Also, it should benefit from longer resident times due to its relatively large size. The only potential issue I see is that it's on the upper end of what's generally considered to be able to pass through the ILM and OLM of the retina, but the literature disagrees on this so I need to do some more reading. It may also be possible to get the Mueller glia to endocytize it and redistribute the protein throughout the retina.

I think what I might do before I even start with ERG testing is to inject fluorescent tagged EYS into healthy zebrafish and then see where it migrates using cryosections. I could do that very quickly and for probably less than $1000. If it works and is able to localize to the photoreceptor outer segments, then it would help me get a better initial estimate of the appropriate dosage. Otherwise it would inform me that I need to try subretinal injections, which would be a bit trickier.

>>9554659
Worth a shot

This thread is getting way too off topic. If there are any janitors reading this, would you please be able to clean up the thread?

>>9593685
The vision problems hit towards the end of undergrad, and I've been living a pretty minimalist lifestyle since I graduated in September, while waiting to find out why I suddenly started going blind. In the mean time I've been making money off of cryptocurrency and consulting fees from some numerical optimization software I wrote. This really is not the way of living that I had envisioned for myself though.

>>9594031
Hey man, if you think you can do a better job than me, then I would love your help. But until then, the current approach of giving up and doing absolutely nothing is not working.

>>9594891
What is a genetic eng? Begone impostor!

>>9565614
You just need to change the mitochondria then everything will change

>>9592830
>>9592095
are you having sex, i.e. inserting the penis into the vagina?

can I see her body or her sweet ass and pussy? maybe breasts? please, she is a girl, I need to see her

>>9554766
Yo this is some real Lorenzo's Oil shit.

>>9594979
Not telling you to give up, neither claiming i could do better as when your life depends on this, you have much more motivation than a random guy protecting his ego and this really isn't my area of expertise- i'm pure math MSc doing PhD in organic chemistry, there's very little i can do to help you except perhaps bringing attention of an expert from my institute.
I'm just stating there are plenty of people like you, and so far none of them succeeded. I wish you success, but you should set a more realistic goal otherwise you're going to be disappointed. What would you think of a chemist suddenly deciding he's going to publish a paper on M-theory? You're in a somewhat similar position- you're trying to get into a multidisciplinary field that's pretty damn complex with weak knowledge of its basis. Just learning will take you a year at the very least, then there's the problem of going from theory to practice, which is notoriously difficult (medicinal biochem has almost as many fake papers as psychology). Even for a world-class researcher your goal is pretty hard to achieve.

>>9595096
Definitely. If op succeeds this story could become a movie.

>>9594979
How's the crowdfunding campaign going along?

>>9554648
yo i have scoliosis can i genrtically engineer my back to not be fucked up?
also can i live forever? i really like the idea of not dying.

>>9554778
https://www.google.com/amp/s/www.express.co.uk/news/world/824000/cancer-CURE-leukaemia-Shares-Oxford-biomedica-HIV-virus-children/amp

>>9595209
Realistic goals are for losers. Go big or go home.

I believe in you OP!

>>9554681
Durrr, must test on animuhls

https://www.youtube.com/watch?v=zvJHq2FJPDM

>>9554750
dont see any cops around

>>9596129
>The parents of six-year-old Emily Whitehead allowed the treatment to be trialled on their leukaemia-stricken child after she was told nothing more could be done and sent for hospice care.

>After just ONE injection she was back to full health in weeks and has been in remission for five years.

>>9596129
>CAR-T is HIV
Well that's quiiiiiite a stretch

Day 16 update:

Still reading about pharmokinetics and whether or not EYS will work for a vitreal administration route. I'm still on the fence about it and need to do even more reading. I'm definitely going to try the fluorescence staining that I mentioned yesterday before I proceed to the ERG tests, though. I may also do side-by-side comparison with a nanoparticle delivery vehicle (if I can find one cheap enough).

Also, I came across something very, very interesting today. It turns out that heparan-sulfate matrix proteins like EYS have some really astounding tissue regeneration properties. Agrin is a very close relative of EYS, and when it was injected into the diseased hearts of mice who had been subjected myocardial infarction, it was found to almost totally regenerate the cardiac tissue within the course of a month after a single dose.

https://www.sciencedaily.com/releases/2017/06/170606095033.htm
(summary, go to sci-hub for the pdf)

This whole class of proteins is heavily involved in promoting tissue growth during normal animal development, but in most species they get downregulated with age. So it may be that EYS ends up as a general treatment for stimulating regeneration in all sorts retinal injuries, and not just RP25. I'll have to test it, but if it works for general purpose retinal regeneration too, that's pretty cool.

>>9595209
Normally I would agree with you, but in this case the therapeutic agent has already been designed for me: I just need to produce it and then get it to where it needs to go. That's a much more approachable task for a non-expert. Plus I taught myself computer science and a lot of math topics, so I don't see why this should be any different. Though if you know of anyone at your institution who could help then I would love to work them.

>>9595966
Temporarily on hold until I iron out the delivery mechanism a little bit more. It would be really bad to post a proposal with a glaring flaw, so I want to make sure everything is perfect.

>>9595209
Normally I would agree with you, but in this case the therapeutic agent has already been designed for me: I just need to produce it and then get it to where it needs to go. That's a much more approachable task for a non-expert. Plus I taught myself computer science and a lot of math topics, so I don't see why this should be any different. Though if you know of anyone at your institution who could help then I would love to work with them.

>>9595966
Temporarily on hold until I iron out the delivery mechanism a little bit more. It would be really bad to post a proposal with a glaring flaw, so I want to make sure everything is perfect.

>>9583013

I literally thought "no it's not" right as I moused over your reply. Fucking gold.

>>9562477
It's been a while since you wrote this but I know you're still in the thread.

If you apply to universities, and in the letter/personal statement you write about your condition and what you are motivated to do I am almost 100% sure a few universities will accept you, if not give you some sort of scholarship. You have a degree in mechanical engineering, that's not history. So you have proven you can learn complex concepts etc. Which is all they really care about.

You should delfinitely consider applying. Apply to dozens, ones known for what you want to do. Try Europe (universities are free) as well.

>>9561660
Buy your own if you have the money. Most of the stuff you're gonna need is pretty cheap. http://www.the-odin.com

>>9598115
OP already discovered this and adjusted his plans accordingly:
>>9587350

>>9597549
OP i plan on seeing if i can help you in future.
I plan on messing around with Julia and simulating the distribution of genetic vectors within a mesh of cells over a given amount of time. I plan on using Turing patterns as a predictor of the outcome of the modification in question (if not a better mathematical modal).
I'm still a newbie but plan on getting into it starting summer, so if you could give me anyway to contact you in future that would be great.

My chan-proof email is: tttt63733@gmail.com
Please provide blog or wherever you plan on posting about this, fund-me so i can shill to biotech students at my uni about the project, and any other info you think adequate to be able to contact you.

Also beware, what may work in mice may not work in humans.

Bump

>>9597771
I've considered it, but again I'm concerned because I have a finite amount of time before I go blind. The average PhD takes about as many years to complete as I have years remaining of eyesight. My hope is that by bypassing academic purgatory and going straight to commercialization, I'll be able to get this protein into human trials faster.

>>9598115
Yeah I had looked at them. How reliable are they compared to someone else like ThermoFisher?

>>9598245
Sounds great, I'll send you an email!

Also, some of the other computer oriented things that I want to do are better analysis of the EYS gene, as well as finite element mass transport studies for pharmokinetics. For the gene analysis, I would like to do a more detailed comparison of the domain structure differences between
a. the four alternative splicings of EYS that are important to the retina
b. proteins closely related to EYS, like perlecan and agrin
c. human EYS and fruit fly/ zebrafish EYS
d. human EYS and any genes in the rabbit genome that might potentially be EYS homologs
That way it might be possible to get a better understanding of all the different functions and interactions EYS has, and also how it's regulated. Most of what it would entail is computing the Levenshtein edit distance between the different genes, finding similar repeating patterns ("homologous domains") in the different genes, and then looking up what those shared patterns do.

I'm not sure if any of those sound interesting, but they would be extremely helpful.

>>9556851
>just like in my greys anatomy

Day 17 update:

*Still* reading about drug delivery. It's really not clear whether a naive vitreal injection approach is going to work or not, and I think the only way to come to a definite answer is to try it out and see what happens. I'm going to do a side-by-side comparison of plain vitreal injection, vitreal injection with nanoparticles, and subretinal injection, and then move forward with the one that works best. Subretinal injection is going to be very tricky though, since zebrafish retinas are tiny.

I'm going to spend another day reading about nanoparticle and subretinal drug delivery before moving on. From there I'm to see whether I can do my preliminary pharmokinetic tests for under $500. If yes, I buy some zebrafish and collect my first data in the next 2-3 weeks before I do crowdfunding. If no, I focus on getting crowdfunded and then do the tests once I have sufficient funds.

I'm going to dialyse the proteins into a phosphate buffered saline solution before injecting them. Then I'm going to cryosection the zebrafish retinas at 12, 24, and 48 hour intervals to look for fluorescent tagged EYS under a microscope. Whichever of the three methods localizes the most EYS in the photoreceptor outer segments wins. Hopefully that's going to be the plain vitreal injections, but we'll see.

>>9599732
You don't need a PhD, a two-year graduate program would do wonders. Not only exposing you to equipment but to knowldge of the faculty as well. I don't think you should be throwing all your eggs into one basket. It's your life though, do what you want.

Going to university doesn't mean that you have to abandon your personal project.

>>9600285
Maybe, but I'll have to think about it some more. It's a really high stakes game that I'm playing here because I only have time to attempt this once.

>>9600286
How much time do you have for this attempt?

OP, how realistic do you think it is to change eye color with CRISPR? If the iris is also immune privileged and isolated from the body, couldn't you inject some CRISPR in there to change a few SNPs to give you blue/green eyes? You only need to change 1-3 base pairs.

>>9599732
Could you send me a link to your crowdfunding campaign via email? Would be interested in donating, as being outside the US and going in the direction of physics/math sadly doesn't enable me to help more than that.
My not-so-personal email is:
omon12257@gmail.com

Good luck on your journey.

>>9601146
the question is how to go about injecting it since its a fixed structure (unlike proto-eye cells in an embryo) and thus might hesitate change of any uniformity.

>>9600751
It's pretty ambitious, but I would like to have this ready for human trials within the next 12 months. The sooner I can administer a treatment, the more vision I will be able to save and the more likely it is to work. If I wait too long then I run the risk of developing secondary vitreoretinal toxicity, and at that point it's basically a lost cause.

>>9601146
No idea. I know there's a really controversial way you can do it by burning away the pigment with lasers, but I'm not sure how you could override the existing pigment with gene therapy alone.

>>9601599
Absolutely, and thank you! Also, depending on how fond you are of PDE's or combinatorics, there may be some crossover.

>>9602228
I'll see if I can help you on the math side. Would be great if I could.

Day 18 update:

It looks like these drug companies have a lot of special tricks for getting their anti-VEGF biologics to penetrate the retina. One of the approaches they use is to spike the delivery vehicle with a little bit of Tween 20 chemical detergent. I think by just straight up copying their delivery vehicle formulations I should be able to get decent results. I still need to read up a little bit more on subretinal injection as a backup plan though.

I think I'm going to spend the next couple days working on the crowdfunding page/ research proposal. Hopefully that will help me collect my thoughts about all the stuff I've learned over the last 2.5 weeks and condense it down into a more specific plan.

>>9602708
Awesome! I think I'm going to make a wiki with a list of open tasks, arranged by discipline, in case anyone else wants to contribute.

>>9603108
>>9603113
Forgot trip.

Day 18 update:

It looks like these drug companies have a lot of special tricks for getting their anti-VEGF biologics to penetrate the retina. One of the approaches they use is to spike the delivery vehicle with a little bit of Tween 20 chemical detergent. I think by just straight up copying their delivery vehicle formulations I should be able to get decent results. I still need to read up a little bit more on subretinal injection as a backup plan though.

I think I'm going to spend the next couple days working on the crowdfunding page/ research proposal. Hopefully that will help me collect my thoughts about all the stuff I've learned over the last 2.5 weeks and condense it down into a more specific plan.

>>9602708
Awesome! I think I'm going to make a wiki with a list of open tasks arranged by discipline, in case anyone else wants to contribute. They'll each have a problem description, background info, and estimated difficulty.

>>9603126
Nice, this could turn into a great citizen science project and become the greatest thing that /sci/ has ever done.

>>9603126
Will you post the link to the wiki here?

>>9554826

Do that and in the best case scenario you would just insert some aminoacids and nucleic acids in your blood stream, that would sooner or later get digested but being bacterial in nature I wouldn't be surprised if there was a inflamatory response of some degree(your body doesn't like having DNA outside of it's context)

You need something that brings that CAS9 inside all the required cells, of course have that CAS9 marked in a way that is not inmedietly destroyed and can be imported into the nucleus and act(and thats without accounting for epigenetic shenaningans that at that momment have that sequence demetilated and unaccesible, you would require several tries)

>>9603126
How long would it take to get the wiki ready, especially the difficulty estimations?

>>9603254
>the greatest thing that /sci/ has ever done.
That's a pretty low bar.

test on a hobo and pay him for his time,after all,who would bleve a hobo that says that a mad scientist tried to turn him into a sort of superhuman?

>>9603888
Sure! I'll just keep my name off of it.

>>9603935
We're doing protein replacement, not gene therapy m80

>>9604396
Unfortunately it might be a couple days. I just had another consulting job come in and I'm kind of bogged down right now between both this project and that one.

>>9603126
I'm down to help. I only got biophysics based training but hopefully I can contribute something.

>>9554648
What's the disease OP?

>>9604811

OP, first time in /sci/ and stumbled on this thread.

I'm a current MD/PhD student and my PhD work is in viral-mediated CRISPR. I have several years of lab experience. Here's a couple of the projects I worked on:

-RNA-based gene therapies for HIV (not all that permanent to your situation)

-A broadly antiviral protein with therapeutic potential (not a replacement of a human protein like you're looking at, but the addition of a protein that had an effect on the innate immune system decreasing susceptibility to viral infections) where part of my work was investigating the best mechanism of getting the protein expressed and where you want it (purification then direct injection of the protein, direct injection of mRNA while inducing it to enter cells for a method of expression that is transient, CRISPR-mediated delivery for permanent expression via a viral vector, CRISPR-mediated genetic engineering of fibroblast-derived stem cells from the patient that would then be injected and constitutively express the desired protein and hopefully integrate into the native tissue)

- Current PhD project dealing with viral delivery of CRISPR for beta thalessemias
All that being said, I think what you're trying to do is extraordinarily difficult, but I took the time to read everything you've said and you seem both motivated and pretty quick to pick all of this up. I'd be interested in chatting with you to talk about where you're heading with all of this while you're still in the early stages. I feel like hopefully I'd be able to give you some insight, point you in the right direction for some specific things, and honestly I'm not opposed to meeting in person if you happen to be somewhat near me and trying to help you through some of the logistics.

Here's an email I just made: tim4cpw123@gmail.com

God speed

Day 19 update:

Sorta busy with this other consulting gig. I read a little bit more about anti-VEGF formulations and also subretinal injections. Subretinal injections should be doable enough, but I'll also need some assistive equipment like a surgical camera and an injection guide (I have too much of a tremor). I ought to be able to mount the needle and camera on a jig driven by anti-backlash ballscrew and knobs for me to adjust. I can call in some favors at the machine shop where I went to school to have it made.

I need to finish writing the proposal and find some decent wiki software. I might try using github's wiki feature.

>>9604829
Awesome, I would really appreciate it! Sorry for the slowness in getting a wiki up.

>>9605172
Suspected RP25/ arCRD. Still waiting on the sequencing results from my parents to confirm.

>>9605322
Sounds excellent! I'll shoot you an email!

>>9605594
What kind of consulting gig? If you don't mind going into detail?

>>9597560
>i just need to produce it and then get it to where it needs to go
Another episode of pharmacodynamics are ez or, we are pretty much curing cancer- there are dozens of therapeutic agents, it's just pharmacodynamics that we need to figure out now!
Good luck anyway, you don't seem to be retarded so at least this will be a learning opportunity for you,

>>9605606
Do take your time with that wiki, last thing you want is a half-assed job. Also, did those researchers, that you emailed a while ago, answer?

>>9605606
Really surprising how much you've learned in this three weeks. It's gone form things I could really help with (gene engineering and such) to something very proteins and kinetics-oriented. Anyway, good luck with that, I'm always a bit skeptical of biohacking activities, but you seem to be serious. Hoping to see that wiki soon. Only thing I could contribute with would be that I reccomend that, if you want to produce a protein you can't get in known vectors, don't bother with cDNA libraries and CRISPR, just design the sequence in silico and synthesize it with some company. Even if the gene comes a bit chopped due to synthesis restrictions, you can assemble everything with known GenEng methods (Gibson Assembly and such). May seem like a lot work, but a lot less than what you proposed doing some days ago.

>>9554648
At the beginning of the thread, I was thinking that you are just another brainlet pop-sci faggot who just read too much of buzzword infested articles on some trashy blog, but as I read more and more, it occured to me that you are damn well into it. Shame I am not able to help in any way, since it's not my pair of shoes, but it's nice seeing other anons junping on the bandwagon. 4chan already managed to produce few madmen, so I won't be surprised if with sufficient enthusiasm and devotion you will actually reach into something. Let your dream keep pushing you forward, and let's hope you (or someone else) will find a cure before it's too late. Godspeed, fellow anon.

Good thread, have a bump

>>9554648
>he is going through with it
the madman

>>9598245
>>9599732
>>9601599
>>9602228
>>9602708
>>9603126
this is wholesome, and i am proud

>>9607378
the absolute madman, we love him!

>>9556851
this

Day 20 update:

Didn't have time to make any progress on this today.

This thread is getting near the bump limit, so I'll make another with the wiki and other informational links in the OP once this one dies.

>>9605934
It's for a company doing microseismic detection for subsurface imaging in the oil and gas industry. They scatter a bunch of geophones all over and then listen for sub-0.1 magnitude earthquakes. Then they end up feeding that data into this geophysics model they built which can estimate subsurface wave velocities and seismic energy.

To localize where the quake actually occurs they have to solve this 4D optimization problem of finding the point with the highest estimated seismic energy. It takes their computational model a while to process each candidate point, so they want to find the location with maximum energy in as few samples as possible.

Pic related is one of the animations I made today. The others are more interesting but they're too big.

>>9606548
Not yet, I should follow up with them.

>>9606712
Thanks, I'll have to look into that assembly method! I was considering doing artificial gene synthesis at first, but I shied away from it because it seemed like it would be a real pain to isolate a cell line that had integrated a copy of the gene without any errors.

>>9606236
>>9607378
>>9607924
>>9608292
>>9608293
Thanks guys, I really appreciate it.

>>9608308
Compiling all your previous updates from this thread is a good idea.

OP you are my hero

>>9608308
Don't forget to link to the new thread from this one.

>>9608350
>>9609354
Will do! Also I'm going to try to go through my emails tonight. I've been super busy lately.

>>9608498
Don't heap praise on me yet. There's still a very real possibility that this might not work.

>>9610250
even if it does fail we might learn something for next time.

>>9610285
this

>>9610250
Don't worry about compiling your previous updates, I already did that and sent the text file to you.

Day 21 update:

Still too busy/ no progress. I want to get back on this tomorrow before I lose momentum.

>>9610285
>>9610324
>next time
Heh

>>9610431
Thanks, I really appreciate it!

>>9610250
No, your motivation is inspiring to me. I guess it's a little deceiving to say you're my hero when you are working on a cure for something.

>>9605562
Can you recommend any math youtubers?

>>9555507
>>9561484

>>9561559
Transhumanism and the possibility of surviving past the Singularity is unironically the only reason I haven't killed myself yet

>>9610646
not you in case some other anon here gets the nuts to try.

>>9610707

Whoever wrote that ironic reactionary pseudointellectual trash would probably have their self-esteem benefit from reading this.

https://pdfs.semanticscholar.org/592f/1c841625684c2364a904c62bf4849bb472f0.pdf

>>9564234

>fucking virology not making healing viruses
>fucking genetics not sharing telekinesis
>fucking toxins blowing up everything

Bump

Day 22 update:

The geophysicists are happy and I think I'm finally over the peak level of busyness for this consulting project, so now back to working on protein replacement.

I finally had time to go through my emails and I'm setting up a git repository for this project. That way I can host files relevant to the project, a dedicated project website, as well as the wiki. I'm going to have that up by tomorrow night, but in the mean time I need decide on a name for the project. I don't really know what to call it, but maybe "Eyes Open Project" so that it's a play on "eyes shut homolog"? What does everyone else think?

>>9611151
I guess that's fair. My hope also is that, if it does work, then it will also make people not be as complacent about attempting to treat other forms of RP. Sure, it's genetic, but so is heart disease.

>>9596439
>Soyboy ruins everything

>>9611306
>Kaczynski
>pseudo-intellectual
almost everything he said would happen has come to pass.

>>9613001
I think that the name "Eyes Open Project" is ok, I certanly can't come up with anything catchier. Promotional text is also important in a crwdfunding campaign so I say you should also think about that.

>>9603120
>>9587350
>>9575459
Bravo OP, I admire your balls and your brain.

If you need crowdfunding, have you heard of this page? it's a web plataform for crowdfunding experiments:

https://experiment.com/

>>9613016
Even if it doesn't, we will still learn something from it. So it's a win/win for us and the global scientific community.

Hey OP, biotech phd here. Really admire your commitment but dont underestimate the production process of proteins. Godspeed

Day 22 update:

The git repo and wiki are up. I'll post them tomorrow in the OP of the new thread once they're a bit more filled in. The list of tasks so far is:

General
-Marketing materials and branding
-Public outreach campaign
-FDA compliance

Bio/Medicine/Chem
-Delivery vehicle formulation
-Protein manufacturing process
-Animal model gene knockout

Math/Physics/CS
-Gene distance calculations/ comparative genomics
-Predicted dose range confidence interval
-Pharmokinetic finite element modelling

Engineering
-Electroretinogram design
-Subretinal injection apparatus design
-Protein manufacturing process

Day 23 update:

The git repo and wiki are up. I'll post them tomorrow in the OP of the new thread once they're a bit more filled in. The list of tasks so far is:

General
-Marketing materials and branding
-Public outreach campaign
-FDA compliance

Bio/Medicine/Chem
-Delivery vehicle formulation
-Protein manufacturing process
-Animal model gene knockout

Math/Physics/CS
-Gene distance calculations/ comparative genomics
-Predicted dose range confidence interval
-Pharmokinetic finite element modelling

Engineering
-Electroretinogram design
-Subretinal injection apparatus design
-Protein manufacturing process

>>9613654
Yeah that's a good point. I've included some outreach tasks on the wiki.

>>9613689
Thanks anon, I'll take a look at it!

>>9614511
I'm going to be purchasing purified EYS from an off the shelf supplier for the first couple experiments. After that I'm hoping to secure enough funding to produce my own, and depending on how much VC money I can raise, also hire some people to work on this project with me.

>>9615546
Literally no-one is going to fund your crap. You'll go blind, your girlfriend will be disappointed and will silently fuck men in front of you knowing you will be none the wiser.

>>9615692
edgy

>>9615538
Will these tasks be divided into subtasks?

>>9615546
Some project names for consideration:
>Operation Occullar
>Project Perception
>EYS wide open
>project EYS
etc

>>9615538
what will be the name of the new OP?

Do it for the meme lad.

>>9615692

>>9616954
>EYS wide open
it's perfect

>>9615538
Can't wait to help with the work in any way I can. I'm rooting for you OP

NEW THREAD
>>9618267
NEW THREAD
>>9618267
NEW THREAD
>>9618267

>Over 23 days
this could be a record.

None of you know what you're talking about. You're all fucking morons. That's why you're here. Don't kid yourselves...you're all fucking morons. I just come here to tell you that. FUCK ALL YOU

>>9619374
>t. neet who's never had the ambition to do anything meaningful