Rapid genome editing by CRISPR-Cas9-POLD3 fusion.
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Authors
Labun, Kornel
Keskitalo, Salla
Soppa, Inkeri
Mamia, Katariina
Tolo, Eero
Meza-Zepeda, Leonardo A
Lorenz, Susanne
Cieslar-Pobuda, Artur
Bordin, Diana L
Valen, Eivind
Publication Date
2021-12-13Journal Title
Elife
ISSN
2050-084X
Publisher
eLife Sciences Publications, Ltd
Volume
10
Language
eng
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Reint, G., Li, Z., Labun, K., Keskitalo, S., Soppa, I., Mamia, K., Tolo, E., et al. (2021). Rapid genome editing by CRISPR-Cas9-POLD3 fusion.. Elife, 10 https://doi.org/10.7554/eLife.75415
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
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.
Keywords
Molecular biology, Cell biology, Gene Editing, Crispr-cas9
Identifiers
34898428, PMC8747517
External DOI: https://doi.org/10.7554/eLife.75415
This record's URL: https://www.repository.cam.ac.uk/handle/1810/333116
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