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A Versatile Viral System for Expression and Depletion of Proteins in Mammalian Cells

Figure 2

Maps of the Entry vectors for either RNAi-mediated protein depletion or protein overexpression.

A) shRNAs or miRNAs can be expressed with either constitutive (H1, U6, CMV) or inducible (H1/TO, CMV/TO) promoters. B) cDNAs encoding the protein of interest (POI) can be cloned into Entry vectors with no tags or different tags for either detection or purification. (Abbreviations: CBP, calcium-binding peptide; SBP, S-binding peptide; GST, glutathione-S-transferase; FKBP12DD, FKBP destabilizing domain. LEFT: Promoter-less Entry vectors. RIGHT: Entry vectors for EF-1α promoter-driven protein expression. In this case, Entry vectors containing a promoter should be recombined with a promoter-less Destination vector (see Figure 4).

Figure 2