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. 2011 May 2;19(5):823–825. doi: 10.1038/mt.2011.70

Figure 2.

Figure 2

Sensor assay to select potent shRNAs. A library of 20,000 short hairpin RNAs (shRNAs) was cloned into an miR30 scaffold under the control of a Tet-inducible promoter (IND) in a vector (pSensor) expressing a green fluorescent protein (GFP) marker gene (Venus) linked to a “sensor” target region containing the cognate 50-nt target region from nine different genes targeted for silencing. Following induction, the shRNAs that are able to inhibit their target will in turn reduce or block GFP expression. The most potent shRNAs can be then isolated following fluorescence-activated cell sorting (FACS) of the cells that lose GFP expression. Nucleotides found at high frequency in the most effective shRNAs are indicated. PGK, phosphoglycerate kinase promoter; RISC, RNA-induced silencing complex; siRNA, small interfering RNA.