FIG 5.

Lentiviral vectors expressing S. pyogenes Cas9 and sgRNAs specific for the HPV-18 E6 and E7 genes induce the death of cervical carcinoma cells. HeLa cells were transduced with a lentiviral vector expressing eGFP, to control for lentiviral toxicity (LCE), or a lentiviral vector expressing S. pyogenes Cas9 and a nonspecific sgRNA or E6- or E7-specific sgRNAs. (A) HeLa cells were transduced with the lentiviral vector at an MOI of ∼2.2, resulting in transduction of ∼90% of the cells in culture. Cell growth was then monitored over a 10-day period. Data from a representative experiment, derived from three similar independent experiments, are presented. (B) HeLa cells were transduced with the lentiviral vector at an MOI of ∼37, which is predicted to transduce >99% of the cells in culture. The cells were then assayed for viability by MTT staining on day 10. Averages of data from two independent experiments normalized to values for a mock-transduced culture are shown. N.S., nonspecific.