how long will it takes to silence target gene after induction of shRNA - (Aug/18/2010 )
Hi everyone,
I've met a problem when assessing the efficiency of transfection. It is supposed that the cells I work with will appear apoptosis after knockdown the target gene. So, will the shRNA silence the gene within 24h? And will the cells die that soon so that I can just observe few fluorescence?
Thanks a lot!
sprina on Thu Aug 19 03:46:32 2010 said:
Hi everyone,
I've met a problem when assessing the efficiency of transfection. It is supposed that the cells I work with will appear apoptosis after knockdown the target gene. So, will the shRNA silence the gene within 24h? And will the cells die that soon so that I can just observe few fluorescence?
Thanks a lot!
Hi there. I would say that normally 24h is too soon. Sometimes you can see the inhibition at arn level at 24-48h, but for protein levels I would wait 48h at least.Anyway you can try and see. Dont you have any resistance in your shrna?
Knocking down some genes causes an apoptotic response mediated by p53. Simultaneous knockdown of p53 has been used to suppress the apoptosis. This is a fairly standard approach using Morpholinos during embryogenesis. It might work with shRNA as well.
Robu ME, Larson JD, Nasevicius A, Beiraghi S, Brenner C, Farber SA, Ekker SC. p53 activation by knockdown technologies. PLoS Genet. 2007 May 25;3(5):e78. Epub 2007 Apr 10.