[Zbrafish] MO rescue guidelines

Giuseppe Borsani via zbrafish%40net.bio.net (by borsani.unibs from gmail.com)
Sun Jun 14 08:19:21 EST 2009


I'm relatively new to zebrafish and, coming from genetics, working on
this animal model is often for me more an art than a science.
We are trying to rescue a MO-induced phenotype through the injection
of capped mRNA. I have a number of questions with apparently no
obvious answers (i.e.: shall I make a construct with or without the 5'
UTR? What about the 3' UTR? Is it convenient to inject a fusion
transcript that includes a reporter gene such as GFP? Since GFP often
alters the functional properties of the gene of interest, what about
bicistronic mRNAs? Shall we inject as a control a mRNA containing a
gene inactivating mutation?

I'm wondering if there is a paper-review-web site with some guidelines
for the strategy to be used.

Thank you for your help

Giuseppe Borsani
University of Brescia



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