using transparent fish for genetic modifications

[Anton Kulaga]

Hi all,

we have some plans about doing genetic modifications of animals in Kiev. As we have all necessary equipment but do not have experience we are going to start with transparent strain of zebra fish called "Casper" ( http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2292119/ ) as:

1) it is very good for learning (to make sure that your are targeting a right encoding gene and that modification really worked) you can first make genetic modifications to make it glow and see if it glows at all and where)

2) we have a project of learning how memory consolidations enzymes work in mind that must be done on living animals (as we want to investigate how learning of the fish correlates with translation of those enzymes), so cameras+computer_vision+transparent_organisms (we have AI experts in team who will do the CS part) looks like a nice solution (just in case if some of you have interest in this project, I put Dmitry Nowicki , the poject leader, in cc to this letter)

So, I wonder: 

1) does anybody of you work with transparent fish or have plans to work (in such case we can investigate dealing with "caspers" together)?

2) Do you know where can we buy it in Europe? I have seen only US websites like http://www.carolina.com/teacher-resources/Interactive/casper-fish-anatomy-studies/tr10794.tr that sell anatomy learning kits but not "fish breeding kits". There is also another problem:  we cannot order the fish via mail to Ukraine, as border control is so cruel and regulations are so stupid there that fish if mailed will likely die or be sent back. That means that we have to transfer it with people who will be traveling to other countries (for instance I am thinking about visiting Poland, Czech and Vienna next month, Dmitry also has some EU plans), so  collaboration with other labs that also want to work with it will be very helpful (we can order it on their address and take some fish by those of us who will happen to travel there).

--
Best regards,
Anton Kulaga

[Mega [Andreas Stuermer]]

 Didn't it save my posts?  :P 

Well, I once was doing some investigation... But as I have no lab with zebrafish experience it didn't develop into a project.

Electroporation/ Lipofetamin transfection of the egg cells should be quite easy. 

 

[Mega [Andreas Stuermer]]

I don't know which ATG is the start codon though. 

http://www.ncbi.nlm.nih.gov/gene/550134

Should be kind of readable from here, but I found no way to show coding site only. 

[Mega [Andreas Stuermer]]

I think I identified the entire promoter + Kozak. I compared the gene to the native ubi. I think translation starts at the underlined sequence. I translated it and did a protein blast on NCBI, looks like ubiquitin.

[Mega [Andreas Stuermer]]

If I am correct, you just have to subsitute the underlined sequence for your Protein of interest.

[Anton Kulaga]

Happy New Year, and thank you, guys, for your suggestions!

2 Dmitry Nowicki, could you provide latest English version of your Rybka Project presentation, so people will know more of what exactly you are going to do with the fish?

2013/12/28 Mega [Andreas Stuermer] <masters...@gmail.com>

If I am correct, you just have to subsitute the underlined sequence for your Protein of interest.

[elegans]

Zebrafish are nice to use because they begin life as a large single-celled embryo. It is very amenable to direct injection of DNA, RNA, or proteins with a glass needle. Ubiquitous promoters are nice to have, but actual learning experiments will require temporal and tissue specific control of the enzymes expression. There are many promoters known for different neuronal cell classes, you'll eventually need to figure out which part of the animal you want to modify.

Here are some helpful startup links: 

http://www.neuro.uoregon.edu/k12/FAQs.html

http://www.jove.com/video/1115/microinjection-of-zebrafish-embryos-to-analyze-gene-function

-Sean

[Mega [Andreas Stuermer]]

A friend told me it is illegal to genetically engineer vertebrates in Austria+Germany other than for biomedical purpose.

While I find this perfectly adequate for mammals, I think the law is overkill for fish. They are not harmed carrying GFP :P

[Nathan McCorkle]

What about jellyfish? That has become a home pet recently!

On Jan 1, 2014 2:56 PM, "Mega [Andreas Stuermer]" <masters...@gmail.com> wrote:

A friend told me it is illegal to genetically engineer vertebrates in Austria+Germany other than for biomedical purpose.

While I find this perfectly adequate for mammals, I think the law is overkill for fish. They are not harmed carrying GFP :P

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[Cathal Garvey]

It's not "illegal", but you need to justify your research. Essentially,
anything with a spine, and also octopus, is regarded in Europe as
deserving of protection from frivolous research or unnecessary suffering.

So, jellyfish does not count, and most invertebrates (all but octopi)
are not covered.

For other animals, I'm not clear on whether it's a legal or conventional
requirement that you get clearance from a regulator or committee, but
it's certainly the norm in Europe. So if you want to do things "by the
book", you'll do fish research in a respectable facility and work under
the oversight of an ethics committee; whether a university, or a
facility and committee of your own construction, following whatever laws
and conventions are expected elsewhere.