To further demonstrate the versatility of sOPTiKD, we generated both hESC and hiPSC lines carrying an inducible shRNA against OCT4 or B2M. Generation of sOPTiKD hPSCs following lipofection was rapid (2 weeks) and extremely efficient, as all the sublines generated showed robust inducible knockdown (Fig. 3B,C). qPCR analyses confirmed that knockdown of OCT4 using sOPTiKD induced differentiation of both hESCs and hiPSCs, whereas knockdown of B2M had no effect (Fig. 3D). Overall, these experiments show that sOPTiKD provides an efficient system to knock down gene expression that can be easily applied to a large number of hPSC lines.
