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Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice
Precise genome editing by systems such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) requires high-efficiency homology-directed repair (HDR). Different technologies have been developed to improve HDR but with limited success. Here, we genera...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978410/ https://www.ncbi.nlm.nih.gov/pubmed/31974493 http://dx.doi.org/10.1038/s42003-020-0768-9 |
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author | Ali, Zahir Shami, Ashwag Sedeek, Khalid Kamel, Radwa Alhabsi, Abdulrahman Tehseen, Muhammad Hassan, Norhan Butt, Haroon Kababji, Ahad Hamdan, Samir M. Mahfouz, Magdy M. |
author_facet | Ali, Zahir Shami, Ashwag Sedeek, Khalid Kamel, Radwa Alhabsi, Abdulrahman Tehseen, Muhammad Hassan, Norhan Butt, Haroon Kababji, Ahad Hamdan, Samir M. Mahfouz, Magdy M. |
author_sort | Ali, Zahir |
collection | PubMed |
description | Precise genome editing by systems such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) requires high-efficiency homology-directed repair (HDR). Different technologies have been developed to improve HDR but with limited success. Here, we generated a fusion between the Cas9 endonuclease and the Agrobacterium VirD2 relaxase (Cas9-VirD2). This chimeric protein combines the functions of Cas9, which produces targeted and specific DNA double-strand breaks (DSBs), and the VirD2 relaxase, which brings the repair template in close proximity to the DSBs, to facilitate HDR. We successfully employed our Cas9-VirD2 system for precise ACETOLACTATE SYNTHASE (OsALS) allele modification to generate herbicide-resistant rice (Oryza sativa) plants, CAROTENOID CLEAVAGE DIOXYGENASE-7 (OsCCD7) to engineer plant architecture, and generate in-frame fusions with the HA epitope at HISTONE DEACETYLASE (OsHDT) locus. The Cas9-VirD2 system expands our ability to improve agriculturally important traits in crops and opens new possibilities for precision genome engineering across diverse eukaryotic species. |
format | Online Article Text |
id | pubmed-6978410 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-69784102020-01-28 Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice Ali, Zahir Shami, Ashwag Sedeek, Khalid Kamel, Radwa Alhabsi, Abdulrahman Tehseen, Muhammad Hassan, Norhan Butt, Haroon Kababji, Ahad Hamdan, Samir M. Mahfouz, Magdy M. Commun Biol Article Precise genome editing by systems such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) requires high-efficiency homology-directed repair (HDR). Different technologies have been developed to improve HDR but with limited success. Here, we generated a fusion between the Cas9 endonuclease and the Agrobacterium VirD2 relaxase (Cas9-VirD2). This chimeric protein combines the functions of Cas9, which produces targeted and specific DNA double-strand breaks (DSBs), and the VirD2 relaxase, which brings the repair template in close proximity to the DSBs, to facilitate HDR. We successfully employed our Cas9-VirD2 system for precise ACETOLACTATE SYNTHASE (OsALS) allele modification to generate herbicide-resistant rice (Oryza sativa) plants, CAROTENOID CLEAVAGE DIOXYGENASE-7 (OsCCD7) to engineer plant architecture, and generate in-frame fusions with the HA epitope at HISTONE DEACETYLASE (OsHDT) locus. The Cas9-VirD2 system expands our ability to improve agriculturally important traits in crops and opens new possibilities for precision genome engineering across diverse eukaryotic species. Nature Publishing Group UK 2020-01-23 /pmc/articles/PMC6978410/ /pubmed/31974493 http://dx.doi.org/10.1038/s42003-020-0768-9 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Ali, Zahir Shami, Ashwag Sedeek, Khalid Kamel, Radwa Alhabsi, Abdulrahman Tehseen, Muhammad Hassan, Norhan Butt, Haroon Kababji, Ahad Hamdan, Samir M. Mahfouz, Magdy M. Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice |
title | Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice |
title_full | Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice |
title_fullStr | Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice |
title_full_unstemmed | Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice |
title_short | Fusion of the Cas9 endonuclease and the VirD2 relaxase facilitates homology-directed repair for precise genome engineering in rice |
title_sort | fusion of the cas9 endonuclease and the vird2 relaxase facilitates homology-directed repair for precise genome engineering in rice |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978410/ https://www.ncbi.nlm.nih.gov/pubmed/31974493 http://dx.doi.org/10.1038/s42003-020-0768-9 |
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