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.
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