Rapid genome editing by CRISPR-Cas9-POLD3 fusion
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...
| Main Authors: | , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2021-12-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/75415 |
| _version_ | 1811203321086607360 |
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| author | Ganna Reint Zhuokun Li Kornel Labun Salla Keskitalo Inkeri Soppa Katariina Mamia Eero Tolo Monika Szymanska Leonardo A Meza-Zepeda Susanne Lorenz Artur Cieslar-Pobuda Xian Hu Diana L Bordin Judith Staerk Eivind Valen Bernhard Schmierer Markku Varjosalo Jussi Taipale Emma Haapaniemi |
| author_facet | Ganna Reint Zhuokun Li Kornel Labun Salla Keskitalo Inkeri Soppa Katariina Mamia Eero Tolo Monika Szymanska Leonardo A Meza-Zepeda Susanne Lorenz Artur Cieslar-Pobuda Xian Hu Diana L Bordin Judith Staerk Eivind Valen Bernhard Schmierer Markku Varjosalo Jussi Taipale Emma Haapaniemi |
| author_sort | Ganna Reint |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T02:53:40Z |
| format | Article |
| id | doaj.art-47f1be8fb06247a18cb89513c9a09958 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:53:40Z |
| publishDate | 2021-12-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-47f1be8fb06247a18cb89513c9a099582022-12-22T03:50:54ZengeLife Sciences Publications LtdeLife2050-084X2021-12-011010.7554/eLife.75415Rapid genome editing by CRISPR-Cas9-POLD3 fusionGanna Reint0https://orcid.org/0000-0003-4823-5485Zhuokun Li1https://orcid.org/0000-0001-7297-6916Kornel Labun2Salla Keskitalo3Inkeri Soppa4Katariina Mamia5Eero Tolo6Monika Szymanska7https://orcid.org/0000-0003-0957-9568Leonardo A Meza-Zepeda8Susanne Lorenz9Artur Cieslar-Pobuda10Xian Hu11https://orcid.org/0000-0002-3381-7514Diana L Bordin12Judith Staerk13https://orcid.org/0000-0001-8698-6998Eivind Valen14Bernhard Schmierer15https://orcid.org/0000-0002-9082-7022Markku Varjosalo16https://orcid.org/0000-0002-1340-9732Jussi Taipale17https://orcid.org/0000-0003-4204-0951Emma Haapaniemi18https://orcid.org/0000-0002-6693-8208Centre for Molecular Medicine Norway, University of Oslo, Oslo, NorwayCentre for Molecular Medicine Norway, University of Oslo, Oslo, NorwayDepartment of Informatics, Computational Biology Unit, University of Bergen, Bergen, NorwayCenter for Biotechnology, University of Helsinki, Helsinki, FinlandCentre for Molecular Medicine Norway, University of Oslo, Oslo, Norway; Stem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Oslo, FinlandCentre for Molecular Medicine Norway, University of Oslo, Oslo, NorwayFaculty of Social Sciences, University of Helsinki, Oslo, FinlandCentre for Molecular Medicine Norway, University of Oslo, Oslo, NorwayDepartment of Core Facilities, Institute for Cancer Research, Oslo University Hospital, Oslo, NorwayDepartment of Core Facilities, Institute for Cancer Research, Oslo University Hospital, Oslo, NorwayCentre for Molecular Medicine Norway, University of Oslo, Oslo, Norway; Department of Cancer Immunology, Institute of Cancer Research, Oslo University Hospital, Oslo, NorwayCentre for Molecular Medicine Norway, University of Oslo, Oslo, NorwayDepartment of Clinical Molecular Biology, Akershus University Hospital, Oslo, NorwayCentre for Molecular Medicine Norway, University of Oslo, Oslo, Norway; Department of Haematology, Oslo University Hospital, Oslo, NorwayDepartment of Informatics, Computational Biology Unit, University of Bergen, Bergen, NorwayDepartment of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, SwedenCenter for Biotechnology, University of Helsinki, Helsinki, FinlandDepartment of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden; Department of Biochemistry, University of Cambridge, Cambridge, United Kingdom; Genome-Scale Biology Program, University of Helsinki, Oslo, NorwayCentre for Molecular Medicine Norway, University of Oslo, Oslo, Norway; Stem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Oslo, Finland; Department of Pediatrics, Oslo University Hospital, Oslo, NorwayPrecision 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.https://elifesciences.org/articles/75415CRISPR-Cas9gene editingmolecular biology |
| spellingShingle | Ganna Reint Zhuokun Li Kornel Labun Salla Keskitalo Inkeri Soppa Katariina Mamia Eero Tolo Monika Szymanska Leonardo A Meza-Zepeda Susanne Lorenz Artur Cieslar-Pobuda Xian Hu Diana L Bordin Judith Staerk Eivind Valen Bernhard Schmierer Markku Varjosalo Jussi Taipale Emma Haapaniemi Rapid genome editing by CRISPR-Cas9-POLD3 fusion eLife CRISPR-Cas9 gene editing molecular biology |
| title | Rapid genome editing by CRISPR-Cas9-POLD3 fusion |
| title_full | Rapid genome editing by CRISPR-Cas9-POLD3 fusion |
| title_fullStr | Rapid genome editing by CRISPR-Cas9-POLD3 fusion |
| title_full_unstemmed | Rapid genome editing by CRISPR-Cas9-POLD3 fusion |
| title_short | Rapid genome editing by CRISPR-Cas9-POLD3 fusion |
| title_sort | rapid genome editing by crispr cas9 pold3 fusion |
| topic | CRISPR-Cas9 gene editing molecular biology |
| url | https://elifesciences.org/articles/75415 |
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