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. 2021 Jan 7;16(1):e0223288. doi: 10.1371/journal.pone.0223288

Fig 1. The targeting of HPV 16E7 gene with CRISPR/Cas9 inhibited cell proliferation and restored Rb protein expression.

Fig 1

(A) HPV 16 +ve (TC1 and CasKi) and HPV -ve (C33A) cell lines were treated with Cas9+ 16E7 specific gRNA or control gRNA (nonspecific) for 72 hours before cell viability was determined by MTT assay. (B) HPV 16 +ve (TC1 and CasKi) and HPV -ve (C33A) cell lines were treated with Cas9+ 16E7 or control gRNAs and allowed to form colonies for two weeks, then the number of colonies was counted. (C) TC1 cells were treated with either Cas9+16E7 or Cas9+ control gRNAs for 72 hours before Rb protein expression was determined by western blot. Jurkat lysate was used as a positive control. Beta-actin was used as loading control. (D) TC1 cells were treated with Cas9+16E7 or control gRNAs for 72 hours, then editing efficiency was determined by T7E1 assay. Two DNA concentrations (500 ng or 800 ng per well, 24-well plate) were tested. All data are presented as mean ± SD. Statistical difference was assessed by ANOVA with post-hoc analysis, *p<0.05, **p<0.01, ***p<0.001.