Effect of MOI on lentiviral shRNA knockdown - (Mar/10/2010 )

Does anyone know if:
- increasing the MOI of shRNA virus infection increases the knockdown effectiveness in target cells?
- infecting target cells multiple time increases knockdown effectiveness?
- after infecting with a shRNA lentivirus carrying a GFP reporter, is the level of GFP expression proportional to the knockdown effectiveness i.e. if all the target cells are GFP positive, will FACS sorting for the top 5% GFP intensity give me a population of cells with better knockdown?

Thanks,

Sam

Does anyone know if:
- increasing the MOI of shRNA virus infection increases the knockdown effectiveness in target cells?
YES, but not always true

- infecting target cells multiple time increases knockdown effectiveness?
YES, but not always true

- after infecting with a shRNA lentivirus carrying a GFP reporter, is the level of GFP expression proportional to the knockdown effectiveness i.e. if all the target cells are GFP positive, will FACS sorting for the top 5% GFP intensity give me a population of cells with better knockdown?
YES, ideally top 5% GFP-cells harbor more copies of lentivirus