I am very interested by the recently developed SHORT-LIVED EGFP mutant
(Clontech) and would like to know if, once synthesis is prevented by
whatever way, the decrease in fluorescence is rapidly clear-cut microscope
I'm thinking of using this d2EGFP to test the activity of various promoters
in human myogenic cells. It seems that various common promoters are somehow
shut down for a few days right at differentation of myogenic cells. Or
could it be a phenomenon of "transition-linked increased mRNA instability"
? (sharing of similar observations, ideas and comments are very welcome!)
We have been slightly fooled til now using EGFP because the protein is so
stable. If the promoter becomes unfunctional and/or the mRNA becomes
unstable for 48 hours, you wouldn't notice it, until you test for the
activity of your other protein of interest. Testing for the activities of
our proteins of interest appears far more tedious than testing for d2EGFP
at the FACS.
So I'd like to hear from investigators that have used this new variant.
With best regards and thanks in advance.
Physiology Dept. / University of Geneva Medical School