[Zbrafish] tdTomato use?

Chi-Bin Chien chi-bin.chien at neuro.utah.edu
Thu Oct 5 11:57:10 EST 2006

Hi everyone,

I'm writing to ask if anyone else has experience with expressing 
tdTomato in zebrafish embryos. This is an mRFP variant from the Tsien 
lab (Shaner et al., 2004) that is supposed to be extremely bright. We 
have had the following odd result.

We cloned tdTomato into pCS2+, which allows either in vitro 
transcription of mRNA, or expression from DNA using the sCMV 
promoter. When we inject DNA at the 1-cell stage, we see nice mosaic 
expression of red fluorescence, as expected. However, when we make 
mRNA and inject it, we get no fluorescence whatsoever. Other pCS2+ 
fluorescent protein constructs work perfectly fine in our hands when 
we inject mRNA.

We would very much like to hear if others have had good (or bad) 
results with tdTomato. On paper it is a great fluorophore, so we 
would like to figure out what's happening.

Chi-Bin Chien
< Dept. Neurobiology and Anatomy |       office: 1-801-585-1701 >
< Univ. Utah Medical Center      |          lab: 1-801-585-1702 >
< 20 North 1900 East, 401 MREB   |          fax: 1-801-581-4233 >
< Salt Lake City, UT  84132      | chi-bin.chien at neuro.utah.edu >
< http://chien.neuro.utah.edu                                   >

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