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loss of GFP expression in stable cell lines


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#1 SF_HK

SF_HK

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Posted 07 June 2021 - 08:25 PM

Hi,

 

I established stable cell lines by gene knockout using lentivirus (GFP expression vector, CMV promoter, Puromycin selection) under optimal antibiotic drug selection. I used viral titre generated by a lab member 3 years ago and stored in -80deg. 

 

The gene was interest was knocked out and determined by western blot.  However after 6-8 passages, the cell are loosing GFP expression but my gene of interest remains knocked-out, as determined by western blot again.  Are these cells good enough to start in vitro studies or do I need to select single colonies again from my earlier stocks?

 

Thank you






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