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Rapid genome editing by CRISPR-Cas9-POLD3 fusion

Published version
Peer-reviewed

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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; FundRef: http://dx.doi.org/10.13039/501100006313


Funder: Norwegian Research Council; FundRef: http://dx.doi.org/10.13039/501100005416


Funder: Knut och Alice Wallenbergs Stiftelse; FundRef: http://dx.doi.org/10.13039/501100004063


Funder: Cancerfonden; FundRef: http://dx.doi.org/10.13039/501100002794


Funder: Instrumentariumin Tiedesäätiö; FundRef: http://dx.doi.org/10.13039/501100008413


Funder: Science for Life Laboratory; FundRef: http://dx.doi.org/10.13039/501100009252


Funder: Academy of Finland; FundRef: http://dx.doi.org/10.13039/501100002341

Keywords

Research Article, Cell Biology, CRISPR-Cas9, gene editing, molecular biology, Other

Journal Title

eLife

Conference Name

Journal ISSN

2050-084X

Volume Title

10

Publisher

eLife Sciences Publications, Ltd
Sponsorship
Ministry of Health and Care Services (279922)