Can i expect a silencing effect from a shRNA Lentiviral plasmid in HEK293T(transiently) too?

Ramya Ambur Sankaranarayanan @Ramya-Ambur-Sankaranarayanan

18 May 2016 1 3K Report

I use pGreenPuro Lentiviral construct. The target mRNA is a Histone demethylase present in HEK293T also.

Also what is the average timepoint i should expect an effect(beyond which the silencing effect might disappear)?

Hello,

In general, if your shRNA is expressed from the vector, you should see the KD, the only problem could be low transfection efficiency, since lentiviral vectors are usually quite big. We use puromycin selection and analyse our cells after 3-4 days of selection (i. e. 4-5 days after transfection).

Also, you may just sort the GFP+ cells and analyse them 3-4 days after.

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