Adding genes to zebrafish embryos, newbie, help plz

Hey everyone, I'm new here.

My name's Vince, I'm trying to add a gene to zebrafish embryos for a science fair project. I'm new to biohacking (like brand new) but I'm pretty familiar with the field of biology, I just get lost when it comes down to the nitty gritty technological side. I don't have the money to spend on microinjection equipment, so I was thinking electrofusion (if that's the correct terminology). I'm buying the gene (likely GFP) from an online supplier and I already have a group of spawning zebrafish. Does anyone know where I can find step by step instructions on how to do this and what equipment I will need (preferably as much DIY equip. as possible)? Even a link to information on the basics of this process so I can educate myself properly would be useful. I know this isn't like baking a cake, but with the right resources, I'm sure I could make myself competent. 

Your guidance is very much appreciated!

Comments

  • OR you can just buy some glofish and explain how they're made and then separate them into different groups and see if a factor you change affects how brightly they glow. This would be doable. What you suggest is, to put it lightly, bloody difficult on a budget to nearly impossible. First off you need more than just the gene. You at the very least need a plasmid, which will not last in the fish for very long. It will likely just be degraded after a bit of replication. If you're lucky it would maybe at best stay in a few cells on the final fish but even then I doubt it. Otherwise you need a vector to get the DNA both into the cell but more importantly into the nucleus and the hard part, getting it to stay by integrating into a safe spot on the fish's chromosomes. Can this be done, yes. Can this be done without proper equiptment which will run you a fair tab, I doubt it. I've been pondering this for years, It's why I built my company, so that one day I could buy the stuff it takes to do this. You'd be better off modifying a plant. They take to modification far far far far far far far easier. Agrobacterium kits would be easier to get, the modification techniques are easier and they've been done many many many many times. Make a plant produce gfp. It's still probably gonna cost you a ton since they don't send you much DNA so you usually have to replicate it which requires equipment. And you need to culture the agrobacterium and modify them. And you have to select the agro colonies that are consistently producing the protein. Then you'd have to infect the plants, see which of those take. If you want to make yourself competent I'd suggest a basic genetics course. Or go to the library and there are multiple books on genetic engineering and modification techniques. I can't remember which ones I read way back when... but there are multiple. If this is something your really want to do you're gonna have to put in the work. It's a lot of work but if you pull it off it would be very impressive. It's a lot of work and can be fairly expensive. Tis the plight of science I'm afraid. That all said it's not impossible, its just hard to get into.
  • Electroporation is what you are thinking about.

    Handful of not particularity spot on stuff in the previous comment, but I'm not going to bother with that. It's basically mostly correct.

    Here is a paper about making yourself some glowing fish action. LINK If you can make it through the methods section without having your brain fall out, you are well on your way. Then you just need to the tools. And you really need to do the microinjection in this case. You can't electroporate this. It won't stick well. Also, you'll just blow up your cells.

    Unfortunately, he is pretty on point with the bit about the learning curve. It's steep. Luckily, you are mistaken about the cake metaphor. It is very much like backing a cake. It's just that your flour costs 300 dollars and your easy bake oven costs another couple thousand...

    That said, once you get the know how and the tools, it really becomes a mater of good experimental design. 

    Good luck and if you have specific questions about any steps, we're always here to help.
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