Need stable GFP expression in IPS and embryonic SCs - (Jan/26/2012 )
I want to have stable GFP expression in my mouse IPSCs and embryonic stem cells and was looking for the method which would result in the greatest efficiency. I have heard that lentiviral packaging vectors may offer the best results. Would I just use a control vector expressing GFP which every manufacturer carries and see if I can generate stable clones. Should I use site directed incorporation of a GFP construct?
Lentiviruses usually don't integrate at a specific site (Adenoviruses can, but that's a different story). Also, you may find silencing of the virus after some time (as early as 3 weeks). I would recommend site-directed recombination (electroporation of a targeting vector), there should be some vectors around for Rosa26 or HPRT locus. Good luck!