Evaluation of Homology-Independent CRISPR-Cas9 Off-Target Assessment Methods.

ABSTRACT: The ability to alter genomes specifically by CRISPR-Cas gene editing has revolutionized biological research, biotechnology, and medicine. Broad therapeutic application of this technology, however, will require thorough preclinical assessment of off-target editing by homology-based prediction coupled with reliable methods for detecting off-target editing. Several off-target site nomination assays exist, but careful comparison is needed to ascertain their relative strengths and weaknesses. In this study, HEK293T cells were treated with Streptococcus pyogenes Cas9 and eight guide RNAs with varying levels of predicted promiscuity in order to compare the performance of three homology-independent off-target nomination methods: the cell-based assay, GUIDE-seq, and the biochemical assays CI

SUBMITTER: Chaudhari HG 

PROVIDER: S-EPMC7757695 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature