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ID 110151
著者
刑部, 祐里子 RIKEN Center for Sustainable Resource Science|Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University 徳島大学 教育研究者総覧 KAKEN研究者をさがす
ワタナベ, タカヒト Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University
スガノ, シゲオ Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University
ウエタ, リサ Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University
イシハラ, リョウスケ Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University
シノザキ, カズオ RIKEN Center for Sustainable Resource Science
刑部, 敬史 Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University 徳島大学 教育研究者総覧 KAKEN研究者をさがす
資料タイプ
学術雑誌論文
抄録
Genome editing using the CRISPR/Cas9 system can be used to modify plant genomes, however, improvements in specificity and applicability are still needed in order for the editing technique to be useful in various plant species. Here, using genome editing mediated by a truncated gRNA (tru-gRNA)/Cas9 combination, we generated new alleles for OST2, a proton pump in Arabidopsis, with no off-target effects. By following expression of Cas9 and the tru-gRNAs, newly generated mutations in CRIPSR/Cas9 transgenic plants were detected with high average mutation rates of up to 32.8% and no off-target effects using constitutive promoter. Reducing nuclear localization signals in Cas9 decreased the mutation rate. In contrast, tru-gRNA Cas9 cassettes driven by meristematic- and reproductive-tissue-specific promoters increased the heritable mutation rate in Arabidopsis, showing that high expression in the germ line can produce bi-allelic mutations. Finally, the new mutant alleles obtained for OST2 exhibited altered stomatal closing in response to environmental conditions. These results suggest further applications in molecular breeding to improve plant function using optimized plant CRISPR/Cas9 systems.
掲載誌名
Scientific Reports
ISSN
20452322
6
開始ページ
26685
発行日
2016-03-26
備考
Copyright © 2016 Authors. This work is licensed under a Creative Commons Attribution 4.0 International License.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license,users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
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出版社版DOI
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フルテキストファイル
言語
eng
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出版社版
部局
生物資源系