Developing oncolytic Herpes simplex virus type 1 through UL39 knockout by CRISPR-Cas9.

ABSTRACT:

Objectives

Oncolytic Herpes simplex virus type 1 (HSV-1) has emerged as a promising strategy for cancer therapy. However, development of novel oncolytic mutants has remained a major challenge owing to low efficiency of conventional genome editing methods. Recently, CRISPR-Cas9 has revolutionized genome editing.

Materials and methods

In this study, we aimed to evaluate the capability of CRISPR-Cas9 to manipulate the UL39 gene to create oncolytic HSV-1. Herein, three sgRNAs were designed against the UL39 gene and transfected into HEK-293 cell line followed by infection with HSV-1 KOS.

Results

After three rounds of plaque purification, several HSV-1 mutants were identified by PCR analysis and sequencing. One of these mutations in which 55 nucleotides were deleted

SUBMITTER: Ebrahimi S 

PROVIDER: S-EPMC7395182 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature