The capacity to knock down genes in a variety of differentiated cells would represent a significant advance over existing systems for inducible gene knockdown. To thoroughly test this possibility, we analyzed the efficacy of the OPTiKD and sOPTiKD platforms to knock down an EGFP transgene in hPSCs differentiated into the three germ layers, as well as in a panel of 13 fully differentiated cell types (Fig. 1A). For both methods, qPCR analyses demonstrated strong and inducible knockdown of EGFP transcripts in all lineages tested (Fig. 4A). Microscopy observations confirmed a robust decrease in EGFP protein expression (Fig. 4B), and flow cytometry showed a decrease in EGFP fluorescence of more than 70% for most lineages (Fig. S4A-G).
