Some how I feel this has come up

So I just set up the new fish tank for both my wife and daughter and they like glowfish. I'm sure a few already see where this is going.

I know the gene is from a jellyfish and injected into the fish egg. Then the offspring of the injected fish has the gene to glow and will glow.

Now the obvious question. Can this be adapted to a mammal such as chicken or other animal. I'm clearly thinking human as well but that time line to see it would take AWHILE.
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  • They've done it already in some mammals. Off the top of my head, I remember sheep, pigs, rabbits, and cats. They've also done it with crop vegetables to use as an indicator of soil quality. Cool stuff.

    I would love to glow in the dark myself. I also would like some chromatophores in case I need to hide from predators.
  • If I remember right, the thing about this is that you have to do the alterations at the embryo stage. Once beyond that (eg birth) it becomes incredibly hard (to put it lightly) to change the genes (because instead of just changing a few cells that will replicated the gene down the line you've got to change thousands, if not millions and billions of cells at once).
  • Making things glow under a blacklight is super easy. you just add the sequence for green fluorescent protein or one of the other color derivatives. There's even a commercial kit to do that to bacteria. Mammals re trickier though. I think it'd end up being more a cost of materials issue rather than a practical one. Since these things tend to be fairly straight forward, just expensive to get in to. 
  • I was toying with the idea of maybe doing it to my crested geckos. I breed them and have essentially a never ending supply. After posting my question about the mammals I did some research and discover the glowing cats and dogs from Korea. Still pretty cool. If I make glowing lizards I may have a money maker There
  • Wow just did a search for the proteins for red and came to the conclusion this is so far beyond my understanding that I'd have better luck learning ancient Egyptian.

    It would still be pretty cool having a few baby glowing crested geckos
  • If you have to add this stuff during embryonic stage that yt-video about chickens grown outside the eggshells in some Japanese school might be of interest.
  • I hadn't seen that one. I'm unsure if it would be needed to be grown outside. A gecko egg is very porous and leathery I'm thinking it could either be injected with a very small needle or maybe even absorbed through the shell.
  • short answer yes. They have glowing puppies and rabbits, they make them glow so that they can easily identify if the animal is a clone or not, because clones are not allowed to mate.
  • If you try to modify an animal you are bound to have to work through it's breeding cycle for every test, and to see if the trait is inherited you have to wait for it's full maturation cycle.

    Might another approach be to modify the natural flora (bacteria) of the animal?

    Simpler organisms, faster to reproduce, easier to test, (no ethical concerns about the bacterias quality of life when something goes wrong)

    Swab the creature and build a profile of the bacteria on it's skin, modify one of more of the strains to glow as required, reinfect the animal.

    I have some (many years ago) experience in microbiology and culturing, it's not hard. I'm not sure about the processes required for genetic manipulation, but i'd assume they have to be easier for a single celled organism in a petri dish than a multicellular embryo in an egg?

  • The maturation of a gecko isn't actually that long so I could have this accomplished quicker then most others. Obviously the fish are much faster.
  • Sorry for the quick response and short winded. I had to post fast last night.

    A lot of the work that was done with with fish and other mammals showed from the offspring of the injected parents.

    Wouldn't the bacteria approach not carry on into offspring? That I feel would be a more expensive route to take if it isn't inherited.

    I feel like your approach would be simpler but is for an already hatched critter vs one in an egg developing.

    Sorry for my poor knowledge on biology it's far from my strong suit.

  • Bacteria changes would be germ line (inherited by offspring), my thought was just that you could try 100 different samples in a day and 100 more the next... even gecko don't breed that fast :)

  • Ok i see what you mean now lol. I'll have to look up some things that you've said so I could see what it actually takes.

    From what I'm gathering I'd be changing the parent geckos bacteria (still not 100% understand that part but that's why they made Google;) ) and the observable change would be in offspring assuming it all took.

    I believe bacteria GFP is actually cheaper then the other option for injecting embryos. I'd have to look again.

    On a side note how feasible is it to harvest the protein from a glow fish and make that usable?
  • I've also seen the glowing rats and strawberries. Instead of rewriting your genes, couldn't it be ingested in some form and programmed to manipulate your cells? That sounds simpler. 
  • I don't honestly know if it would be.

    I'm hoping some one with more biology back ground could chime in with some 2 cents on how it could even be accomplished.

    @dr_allcome do you know what it would require to simply get to the point of being capable of doing this with the bacteria? I mean equipment wise.
  • So here's the thing. Making things glow is really easy, though it depends what you mean by glow. Those strawberries and rats etc don't just glow (usually). They've *usually* been modified to have GFP (green flourescent protein) as I said before. GFP is super easy to work with and is used often as a reporter. They stick the sequence for gfp directly next to the thing they want the organism to express and so if the cells glow, it's proabbly making the other thing you want. Thing is though, it's green FLOURESCENT protein. This almost always means it glows under a black light. There are other variations of the protein that will give different colors.

    If you want to make bacteria glow there are plasmids you can buy, the one I'm familair with is the pglo system. LINK to a whole thing explaining exactly how to use it. Basically they've put the code for GFP onto a plasmid that also encodes ampicilin resistance. You use either calcium or electroporation or some other technique to get the plasmid into your sample of bacteria then you plate them onto agar plates that contain ampicilin. In theory the only bacteria that can grow are the ones that contain the glowing protein. This isn't actually true as some will randomly mutate or be resistant and will grow without glowing, but you can easily isolate the glowing bacteria from the non glowing ones, take a sample, grow them in batch and get a bucket worth of glowing bacteria. In theory you could do this to any bacteria, potentially even one that naturally makes up skin flora. So if you inoculate an organism with the bactera their fur/skin/whatever would get covered partially in the bacteria and they would apear to glow. How you would control this so it doesn't get everywhere and you don't end up glowing, I dunno. 

    Doing the same thing to an animal is much harder, 1 because the plasmid you need is much more complicated and as a rule, noobs should never work with animal cells. Compared to bacteria they are a giant paint in the ass to work with. That said, you could probably build a gene gun, get some frog/fish eggs and just blast the DNA you want in and hope for the best. (no one said biology was boring, we get guns too) Nice thing is that way even if 100 eggs don't work, maybe 1 will. In fact, heres a paper talking about exactly that LINK. If nothing else it'd be fun to build a gene gun and try this out. 

    Now back to the glowing. As I said before, most of the things you've seen are fluorescent due to illumination by a black light. As you know from fireflies and other glowing organisms, they glow just fine on their own. This is because they work ona  different principle (though technically gfp can be used for this but more on that later). What that's technically called is autofluorescence. It works on an enzyme-substrate system. In fireflies this luciferin (substrate) and luciferase (enzyme). Soon as the two mix, they make light. In jelly fish GFP actually does the same sort of thing but with a different enzyme. There's a group using the firefly system to make plants glow LINK. To do this to an animal you'd need to encode all of the enzymes needed to make the substrate luciferin and of course the luciferase. You'd need to play with how well all these things are expressed as it can be difficult to get thigns at the right levels to really glow brightly all the time. One of the reasons the glowing plant people are taking so long to deliver. 

    @EmberErikson That's actually a lot harder. You'd need a viral vector to do that which has it own host of issues and also all of the cancer is a legitimate issue. If you just want to glow temporarily, I was talking about it in another thread, you could use something like a liposome to deliver the plasmid to your cells in a local area. So long as it doesn't try an integrate and doesn't just get chewed up by your immune system it should cause the cells affected to express the protein until they die. When they do the plasmid will be destroyed and you'll stop glowing. I'd stick with something simple like gfp. It's still difficult but less than a virus. Again, own host of issues, but that's a topic for another day. LINK to other thread

    Also, if ya'll want to learn all this kind of stuff, might i suggest exosphere? LINK
  • @glims lol I posted the same link at the same time 
  • That was a lot of info to digest. I will probably be reading that a few times (hopefully after a full nights sleep).

    Thank you both for that massive amount of info and I'll definitely look at exosphere.
  • I got home from work and saw a note for myself to post some links for basic microbiology equipment... kinda redundant and my 15 year ago rusty lab skills are way outclassed by you guys.

    Soooo happy I found this forum :) 
  • @chironex I'm gonna tackle one thing at a time for my own sanity with this.

    Pglo:
    With that kit it would alow me to strictly modify bacteria? This would in fact alow me to do as @dr_allcome mentioned and infect the animal (lizard) with the affected bacteria (hopefully) not harming it in the process assuming I used bacteria from the lizard (natural flora)?

    Glowing:
    Fully understand the glowing under black Light. Many geckos are often kept under black Light in general at night because it doesn't affect their vision negatively. So a black Light based glow is acceptable.

    Guns:
    Would this be needed (never thought I'd say that under the word guns) if you were to go the "roundabout" rt with bacteria still?

    Exosphere:
    Some how I missed your video on it and the last link you posted to it was back to this thread. I did get to the site though. Unfortunately I am stuck on the east coast without the ability to travel and spend that amount of time out of work. :(

    I feel like I missed something but I think this covers my foggy brain thoughts. I'll have to look at the posts again after some sleep.

    Side note the most biology I know is Pennetta squares which I used mostly for breeding geckos. I'll try and keep up with what's posted on this very weak subject of mine.
  • As I mentioned, how do you control the spread of that bacteria? if you release it how do you stop it gettig on everything the gecko touches? If it got around that plasmid could be picked up by other bacteria and they would also get ampiciliin resistance which would also come with increased resistance to other antibiotics. This isn't really the end of the world but it's one reason to be careful with gmo organisms. Point is, no that's not really a viable idea. If you want a glowing animal, modify it not it's bacteria. But making some glowing bacteria is a good way to learn the basics of genetic engineering. So yes, you'd need a gene gun or something similair to modify an animal. Though if you wanted to make glowing plants there are bacteria that inject their DNA into plants. So you can modify the bacteria, then infect a plant with it to transfer the DNA.
  • Ok I missed the spread the bacteria part. I should definitely move slow in this sense if I do go and try to do anything remotely like this. I am definitely a noob in this area and as you said I should stay clear of my animals for now.

    I suppose the next thing I should probably do it learn about a gene gun and the simpler genetic engineering. That kit is a very welcoming price to get though which is nice.
  • I'd be ok with glowing temporarily. It'd kind of hard to explain why i was glowing when I walk into a dark room. Awesome conversation starter though! 
    And I think that cancer will always be a concern in any case though I'll admit that I don't know biology very well.
  • @embererikson I think the kicker was resistant to antibiotic bacteria.

    I did a look over on the gene guns and it seems very simplistic in design. I'm hoping I'm wrong because I'm sure it costs a good amount of money to buy one.

    1)force builds up behind disk
    2)disc breaks and air flow travels through "barrel"
    3)hits nano particles/DNA?
    4)force pushes particles into cells of plant/animal

    A wiki searched indicated that an air soft gun was originally used and then refined.....
  • theyre expensive to buy but i've come to learn that that doesn't mean things are hard to make. Here's the first thing that came up on google when i search DIY gene gun LINK. Im sure there are better, but point is, science doesn't have to be expensive. You just need to be clever to offset it. If you want to mess with gene guns, start with plants. They're reasonably forgiving all things considered. Just work with onions or something. See if you can make em glow. Obvs there's a fair bit of reading you'll need to do but if you're gonna chase this (which i HIGHLY recommend cause frankly it's awesome) I'd be happy to point you in the right direction
  • edited June 2016
    Absolutely I would love to mess with this. Anything you can do to point or steer me in any sort of direction will help. I just have a hard time doing projects because I'm kinda between jobs which really sucks the money out of me. (Seasonal again) hopefully my new job comes through and I'll get a regular income which would make this sort of thing and others that interest me more and more possible.

    I've been looking at glass ware and such to try and start up having equipment to do more then just electronics.

    EDIT: forgot to mention I was just watching a video of Pic2 being made and was going to do that route for making one. I just wasn't sure it would function. Again it seemed to simple.
  • Duckweed would be another good plant to start with, simple and grows fast. It's also tiny and basically clone propagates itself. 
  • To start off maybe check out Biopunk:solving the worlds biggest problems in kitchens and garages. I just started reading it and it's pretty damn good so far.

    For a more technical book here's this LINK

    I'd absolutely recommend the biology video series by crash course LINK

    Also if you haven't seen it before, one of the best series on youtube LINK. They cover everything from biology to chemistry to physics. Start watching and be prepared to lose a few hours when you can't stop.

  • that's a badass channel. thanks @chironex , always nice to learn something new :)
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