Improving CRISPR-Cas9 directed faithful transgene integration outcomes by reducing unwanted random DNA integration.

ABSTRACT:

Background

The field of genome editing has been revolutionized by the development of an easily programmable editing tool, the CRISPR-Cas9. Despite its promise, off-target activity of Cas9 posed a great disadvantage for genome editing purposes by causing DNA double strand breaks at off-target locations and causing unwanted editing outcomes. Furthermore, for gene integration applications, which introduce transgene sequences, integration of transgenes to off-target sites could be harmful, hard to detect, and reduce faithful genome editing efficiency.

Method

Here we report the development of a multicolour fluorescence assay for studying CRISPR-Cas9-directed gene integration at an endogenous locus in human cell lines. We examine genetic integration of reporter genes in transientl

SUBMITTER: Hermantara R 

PROVIDER: S-EPMC10964699 | biostudies-literature | 2024 Mar

REPOSITORIES: biostudies-literature