Abstract:
High risk human papilloma viruses are the main etiological agents of cervical cancers
responsible for approximately 99.7% of all cases. The persistence of viral infection is the major
reason that triggers oncogenesis in host’s infected cells due to the deregulation of oncogenes
E6 and E7. The current treatment regimens include cisplatin base chemotherapy (CRT),
radiotherapy and hysterectomy, however, these approaches have their downfalls as CRT has a
recurrence rate of 25-40% and surgery results in loss of fertility. CRISPR-Cas9 therapy can
prove to be an effective approach to curb viral persistence targeting HPV oncogenes at genetic
level and can be an effective remedy for cervical cancer. This study employs the use of
CRISPR-Cas9 gene editing system to target E6 and E7 genes of HPV-18 virus. Novel guide
RNAs against oncogenes E6 and E7 genes of HPV-18 were designed and their off-targets in
human genome were evaluated of CasOFFinder software. They were then cloned in pX260
CRISPR Cas 9 expression vector. The confirmation of cloning was done by sangar sequencing.
The resultant E6 gRNA-CRISPR Cas9 and E7 gRNA-CRISPR Cas9 vectors can be potentially
therapeutic against HPV-18 after their efficacy is evaluated in vitro and in vivo.
