[Zbrafish] Re: Zebrafish transgenics

Jean-Pierre Levraud via zbrafish%40net.bio.net (by jlevraud from pasteur.fr)
Thu Sep 27 06:02:29 EST 2007


Hi Tania,
if what you want is simply to overexpress a protein, there is a very
simple
way to overcome mosaicism, which is not familiar to mice people:
instead of
injecting DNA, you can inject 1-cell embryos with in vitro transcribed
mRNA.
This results in ubiquitous expression and is very efficient (but
expression lasts
only for a few days).
I suggest you have a look at this paper:
Hyatt & Ekker, "Vectors and techniques for ectopic gene expression in
zebrafish",
Meth Cell Biol, 59:117 (1999)
regards,
:@)
Jean-Pierre


On Sep 25, 8:18 pm, "Tania Vinagre" <vinagre.ta... from gmail.com> wrote:
> I work with mice and consequently I have some limitations regarding numbers
> of transgenic embryos I can obtain. For this reason, I am considering doing
> some experiments in zebrafish. I am interested in getting a reasonably high
> number of transgenic embryos and analyzing their skeletal phenotype. At the
> moment I am not interested in producing stable lines, only embryos, but I
> came across a number of possible problems. Is there any simple way to
> overcome the mosaicism in zebrafish transgenics? How can I promote the
> integration of my construct? Is there any good review on transgenic
> techniques?
>
> Thanks in advance



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