Rapid genome editing by CRISPR-Cas9-POLD3 fusion

Authors
Labun, Kornel 
Keskitalo, Salla 
Soppa, Inkeri 

Change log
Abstract

Precision CRISPR gene editing relies on the cellular homology-directed DNA repair (HDR) to introduce custom DNA sequences to target sites. The HDR editing efficiency varies between cell types and genomic sites, and the sources of this variation are incompletely understood. Here, we have studied the effect of 450 DNA repair protein-Cas9 fusions on CRISPR genome editing outcomes. We find the majority of fusions to improve precision genome editing only modestly in a locus- and cell-type specific manner. We identify Cas9-POLD3 fusion that enhances editing by speeding up the initiation of DNA repair. We conclude that while DNA repair protein fusions to Cas9 can improve HDR CRISPR editing, most need to be optimized to the cell type and genomic site, highlighting the diversity of factors contributing to locus-specific genome editing outcomes.

Publication Date
2021-12-13
Online Publication Date
Acceptance Date
2021-11-15
Keywords
Research Article, Cell Biology, CRISPR-Cas9, gene editing, molecular biology, Other
Journal Title
eLife
Journal ISSN
2050-084X
Volume Title
10
Publisher
eLife Sciences Publications, Ltd
Sponsorship
Ministry of Health and Care Services (279922)