Gene knock-out chain reaction enables high disruption efficiency of HPV18 E6/E7 genes in cervical cancer cells

• DOI: 10.1016/j.omto.2021.12.011
• Publication Date: 2021-12-18T03:51:58Z
• Authors: Rui Tian; Jiashuo Liu; Weiwen Fan; Rui Li; Zifeng Cui; Zhuang Jin; Zhaoyue Huang; Hongxian Xie; Lifang Li; Zheying Huang; Zheng Hu; Ping Zhou; Xun Tian
• Keywords: Gene knockin

A genome editing tool targeting the high-risk human papillomavirus (HPV) oncogene is a promising therapeutic strategy to treat HPV-related cervical cancer. To improve gene knockout efficiency, we developed chain reaction (GKCR) method for continually generating mutagenic disruptions and used this disrupt HPV18 E6 E7 genes. We verified that GKCR Cas9/guide RNA (gRNA) cassettes could integrated into targeted loci via homology-independent insertion (HITI). The qPCR results revealed enabled relatively higher Cas9/gRNA cassette rate than control (the common CRISPR-Cas9 strategy). Tracking of Indels by DEcomposition (TIDE) assay showed produced significantly percentage insertions or deletions (indels) in Furthermore, E6/E7 oncogenes, found upregulated P53/RB proteins inhibited proliferation motility HeLa cells. improved efficiency which might provide new insights treatment HPV infection related