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Precise and heritable gene targeting in rice using a sequential transformation strategy
Gene targeting (GT) is a powerful tool for modifying endogenous genomic sequences of interest, such as sequence replacement and gene knockin. Although the efficiency of GT is extremely low in higher plants, engineered sequence-specific nucleases (SSNs)-mediated double-strand breaks (DSBs) can improv...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9939429/ https://www.ncbi.nlm.nih.gov/pubmed/36814841 http://dx.doi.org/10.1016/j.crmeth.2022.100389 |
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author | Zhang, Wenxin Wang, Rui Kong, Dali Peng, Fangnan Chen, Mei Zeng, Wenjie Giaume, Francesca He, Sheng Zhang, Hui Wang, Zhen Kyozuka, Junko Zhu, Jian-Kang Fornara, Fabio Miki, Daisuke |
author_facet | Zhang, Wenxin Wang, Rui Kong, Dali Peng, Fangnan Chen, Mei Zeng, Wenjie Giaume, Francesca He, Sheng Zhang, Hui Wang, Zhen Kyozuka, Junko Zhu, Jian-Kang Fornara, Fabio Miki, Daisuke |
author_sort | Zhang, Wenxin |
collection | PubMed |
description | Gene targeting (GT) is a powerful tool for modifying endogenous genomic sequences of interest, such as sequence replacement and gene knockin. Although the efficiency of GT is extremely low in higher plants, engineered sequence-specific nucleases (SSNs)-mediated double-strand breaks (DSBs) can improve GT frequency. We recently reported a CRISPR-Cas9-mediated approach for heritable GT in Arabidopsis, called the “sequential transformation” strategy. For efficient establishment of GT via the sequential transformation method, strong Cas9 activity and robust DSBs are required in the plant cells being infected with Agrobacterium carrying sgRNA and donor DNA. Accordingly, we generated two independent parental lines with maize Ubiquitin 1 promoter-driven Cas9 and established sequential transformation-mediated GT in the Japonica rice cultivar Oryza sativa Nipponbare. We achieved precise GFP knockin into the endogenous OsFTL1 and OsROS1a loci. We believe that our GT technology could be widely utilized in rice research and breeding applications. |
format | Online Article Text |
id | pubmed-9939429 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-99394292023-02-21 Precise and heritable gene targeting in rice using a sequential transformation strategy Zhang, Wenxin Wang, Rui Kong, Dali Peng, Fangnan Chen, Mei Zeng, Wenjie Giaume, Francesca He, Sheng Zhang, Hui Wang, Zhen Kyozuka, Junko Zhu, Jian-Kang Fornara, Fabio Miki, Daisuke Cell Rep Methods Article Gene targeting (GT) is a powerful tool for modifying endogenous genomic sequences of interest, such as sequence replacement and gene knockin. Although the efficiency of GT is extremely low in higher plants, engineered sequence-specific nucleases (SSNs)-mediated double-strand breaks (DSBs) can improve GT frequency. We recently reported a CRISPR-Cas9-mediated approach for heritable GT in Arabidopsis, called the “sequential transformation” strategy. For efficient establishment of GT via the sequential transformation method, strong Cas9 activity and robust DSBs are required in the plant cells being infected with Agrobacterium carrying sgRNA and donor DNA. Accordingly, we generated two independent parental lines with maize Ubiquitin 1 promoter-driven Cas9 and established sequential transformation-mediated GT in the Japonica rice cultivar Oryza sativa Nipponbare. We achieved precise GFP knockin into the endogenous OsFTL1 and OsROS1a loci. We believe that our GT technology could be widely utilized in rice research and breeding applications. Elsevier 2023-01-17 /pmc/articles/PMC9939429/ /pubmed/36814841 http://dx.doi.org/10.1016/j.crmeth.2022.100389 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Article Zhang, Wenxin Wang, Rui Kong, Dali Peng, Fangnan Chen, Mei Zeng, Wenjie Giaume, Francesca He, Sheng Zhang, Hui Wang, Zhen Kyozuka, Junko Zhu, Jian-Kang Fornara, Fabio Miki, Daisuke Precise and heritable gene targeting in rice using a sequential transformation strategy |
title | Precise and heritable gene targeting in rice using a sequential transformation strategy |
title_full | Precise and heritable gene targeting in rice using a sequential transformation strategy |
title_fullStr | Precise and heritable gene targeting in rice using a sequential transformation strategy |
title_full_unstemmed | Precise and heritable gene targeting in rice using a sequential transformation strategy |
title_short | Precise and heritable gene targeting in rice using a sequential transformation strategy |
title_sort | precise and heritable gene targeting in rice using a sequential transformation strategy |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9939429/ https://www.ncbi.nlm.nih.gov/pubmed/36814841 http://dx.doi.org/10.1016/j.crmeth.2022.100389 |
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