plasmid transfection and virus infection simutaneously - (Jul/31/2005 )

Dear All,

One of my interested gene is expressed in lentivirus vector, the other is plasmid DNA. I want to get both into cells and want high transfection efficiency. What should I do? Should I do plasmid transfection first and then after several hours (5-8hrs) infect the target cells again with virus? Or do the virus infection first then come with plasmid transfection?

Has anybody have such experience to share or any suggestions? Thanks a lot! Really appreciate it.

apple.

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why not cotransfect both together? both are vectors anyway, but how good expressio you get from lentiviral vector? i use retroviral vectors for simple transfections, and get decent expression.
if that's not an option, i would infect first. with a good titer you'll get cells stably expressing one of yours proteins. then, when they recover just transfect.

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Thanks a lot Jusu! I got more than 90% transfection efficiency when infected with lentivirus expressing my DNA, checked by GFP since the vector is double-cassette), but never tried cotransfection of these two vectors. But like you said, I might try to infect cells first and after 48 hours, when cells revocer I will do the transfection. Will let you know how it'll go!
Apple.

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