I can package lentivirus using pGIPZ vector for silence RNA in 293T cells but no virus (?) or the virus doesn't work when I use plenti6.2 vector for overexpression. Same packaging cells, same packaging plasmids (psPAX2, pMD2) and I did it at the same time. Can anyone give me suggestions what can be the problem? Thank you very much. Jen
It would be worth consider which gene targets you are silencing - it might harm the 293T cells
Questions: What is your marker (fluorescent protein, drug selection etc) in pLenti6.2? If it's GFP, can you see expression after transfection? How are you titering your virus? Have you sequenced or digested the plasmid to make sure it's correct?
Jill pointed out most of the concerns above. I think you must test your construct and virus productivity. The other possibility could be the infectivity, most times I use some reagent for higher adsorption such as polybrene or transdux. Hope all these help you sort out the probable cause.
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