ID 110151
Author
Osakabe, Yuriko RIKEN Center for Sustainable Resource Science|Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University Tokushima University Educator and Researcher Directory KAKEN Search Researchers
Watanabe, Takahito Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University
Sugano, Shigeo S Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University
Ueta, Risa Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University
Ishihara, Ryosuke Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University
Shinozaki, Kazuo RIKEN Center for Sustainable Resource Science
Osakabe, Keishi Center for Collaboration among Agriculture, Industry and Commerce, Tokushima University|Faculty of Bioscience and Bioindustry, Tokushima University Tokushima University Educator and Researcher Directory KAKEN Search Researchers
Content Type
Journal Article
Description
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.
Journal Title
Scientific Reports
ISSN
20452322
Volume
6
Start Page
26685
Published Date
2016-03-26
Remark
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/
EDB ID
DOI (Published Version)
URL ( Publisher's Version )
FullText File
language
eng
TextVersion
Publisher
departments
Bioscience and Bioindustry