Integration of high-risk HPV genomes into cellular
chromatin has been confirmed to promote cervical
carcinogenesis, with HPV16 being the most prevalent high-risk type. Herein, we evaluated the
therapeutic effect of the CRISPR/Cas9 system in cervical
carcinogenesis, especially for cervical precancerous lesions. In
cervical cancer/pre-
cancer cell lines, we transfected the HPV16 E7 targeted CRISPR/Cas9,
TALEN, ZFN plasmids, respectively. Compared to previous established ZFN and
TALEN systems, CRISPR/Cas9 has shown comparable efficiency and specificity in inhibiting cell growth and colony formation and inducing apoptosis in
cervical cancer/pre-
cancer cell lines, which seemed to be more pronounced in the S12 cell line derived from the low-grade cervical lesion. Furthermore, in xenograft formation assays, CRISPR/Cas9 inhibited
tumor formation of the S12 cell line in vivo and affected the corresponding
protein expression. In the K14-HPV16 transgenic mice model of HPV-driven spontaneous cervical
carcinogenesis, cervical application of CRISPR/Cas9 treatment caused mutations of the E7 gene and restored the expression of RB, E2F1, and CDK2, thereby reversing the cervical
carcinogenesis phenotype. In this study, we have demonstrated that CRISPR/Cas9 targeting HPV16 E7 could effectively revert the HPV-related cervical
carcinogenesis in vitro, as well as in K14-HPV16 transgenic mice, which has shown great potential in clinical treatment for cervical precancerous lesions.