Protocol for Genome Editing to Produce Multiple Mutants in Wheat

Here, we describe a protocol for producing multiple recessive mutants via genome editing in hexaploid wheat (Triticum aestivum) cv. Fielder. Using Agrobacterium-delivered CRISPR/Cas9 and three sub-genome-specific primer sets, all possible combinations of single, double, and triple transgene-free mut...

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Autores principales: Abe, Fumitaka, Ishida, Yuji, Hisano, Hiroshi, Endo, Masaki, Komari, Toshihiko, Toki, Seiichi, Sato, Kazuhiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580089/
https://www.ncbi.nlm.nih.gov/pubmed/33111098
http://dx.doi.org/10.1016/j.xpro.2020.100053
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author Abe, Fumitaka
Ishida, Yuji
Hisano, Hiroshi
Endo, Masaki
Komari, Toshihiko
Toki, Seiichi
Sato, Kazuhiro
author_facet Abe, Fumitaka
Ishida, Yuji
Hisano, Hiroshi
Endo, Masaki
Komari, Toshihiko
Toki, Seiichi
Sato, Kazuhiro
author_sort Abe, Fumitaka
collection PubMed
description Here, we describe a protocol for producing multiple recessive mutants via genome editing in hexaploid wheat (Triticum aestivum) cv. Fielder. Using Agrobacterium-delivered CRISPR/Cas9 and three sub-genome-specific primer sets, all possible combinations of single, double, and triple transgene-free mutants can be generated. The technique for acceleration of generation advancement with embryo culture reduces time for mutant production. The mutants produced by this protocol can be used for the analysis of gene function and crop improvement. For complete details on the use and execution of this protocol, please refer to Abe et al. (2019).
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spelling pubmed-75800892020-10-26 Protocol for Genome Editing to Produce Multiple Mutants in Wheat Abe, Fumitaka Ishida, Yuji Hisano, Hiroshi Endo, Masaki Komari, Toshihiko Toki, Seiichi Sato, Kazuhiro STAR Protoc Protocol Here, we describe a protocol for producing multiple recessive mutants via genome editing in hexaploid wheat (Triticum aestivum) cv. Fielder. Using Agrobacterium-delivered CRISPR/Cas9 and three sub-genome-specific primer sets, all possible combinations of single, double, and triple transgene-free mutants can be generated. The technique for acceleration of generation advancement with embryo culture reduces time for mutant production. The mutants produced by this protocol can be used for the analysis of gene function and crop improvement. For complete details on the use and execution of this protocol, please refer to Abe et al. (2019). Elsevier 2020-06-13 /pmc/articles/PMC7580089/ /pubmed/33111098 http://dx.doi.org/10.1016/j.xpro.2020.100053 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Abe, Fumitaka
Ishida, Yuji
Hisano, Hiroshi
Endo, Masaki
Komari, Toshihiko
Toki, Seiichi
Sato, Kazuhiro
Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_full Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_fullStr Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_full_unstemmed Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_short Protocol for Genome Editing to Produce Multiple Mutants in Wheat
title_sort protocol for genome editing to produce multiple mutants in wheat
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7580089/
https://www.ncbi.nlm.nih.gov/pubmed/33111098
http://dx.doi.org/10.1016/j.xpro.2020.100053
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