[Zbrafish] Red fluorescent reporters for transgenic fish

Chi-Bin Chien via zbrafish%40net.bio.net (by chi-bin.chien from neuro.utah.edu)
Wed Nov 4 13:51:12 EST 2009


Dear Wei,

Very likely your problem is due to the slow maturation of DsRed.  
mCherry should be better and is fairly bright. We have also had good  
success with TagRFP, which is extremely bright (available commercially  
from Evrogen). Another possibility is tdTomato, which is extremely  
bright. We have not used it ourselves but have heard good things from  
others. Don't know about maturation time.

There is an easy experiment to test whether maturation time is the  
issue: do a DsRed in situ on your transgenics, and see if the  
timecourse mimics the native gene.

best wishes,
Chi-Bin Chien

On Nov 2, 2009, at 5:19 PM, Wei Xia wrote:

> Dear All,
> We are wondering about what kinds of red fluorescent reporters are  
> good for making transgenic zebrafish.
> Our gene’s strongest expression occurs around 24hpf by in situ  
> results. But when we used DsRed as the reporter we could not see any  
> fluorescence until after 44-48hpf in our transgenic embryos. The  
> fluorescence was pretty obvious at 24hpf if transient expression was  
> checked by plasmid injection. So we guess the late detection of  
> DsRed in these transgenic embryos is not because of the promoter we  
> used is not complete or something. We thought this is might because  
> of DsRed’s long maturation half time. We are thinking about using  
> monomeric mCherry but worry about its low brightness compared to  
> DsRed.
> If you have any experiences of making RFP transgenic fish please  
> share them with us!
> Thanks in advance for your testimonies on this.
> Wei
>
> <ATT00001.txt>

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