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Rapid genome editing by CRISPR-Cas9-POLD3 fusion.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Labun, Kornel 
Keskitalo, Salla 
Soppa, Inkeri 

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.

Description

Funder: Barncancerfonden


Funder: Cancerfonden


Funder: Academy of Finland


Funder: Science for Life Laboratory


Funder: Knut och Alice Wallenbergs Stiftelse


Funder: Instrumentariumin Tiedesäätiö


Funder: Norwegian Research Council

Keywords

CRISPR-Cas9, cell biology, gene editing, molecular biology, CRISPR-Associated Protein 9, CRISPR-Cas Systems, Cells, Cultured, DNA Polymerase III, DNA Repair, Gene Editing, Humans

Journal Title

Elife

Conference Name

Journal ISSN

2050-084X
2050-084X

Volume Title

10

Publisher

eLife Sciences Publications, Ltd