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ID 114487
タイトル別表記
Multiplex Genome Editing in Tomato
著者
Hashimoto, Ryosuke Tokushima University
Ueta, Risa Tokushima University
Abe, Chihiro Tokushima University
キーワード
CRISPR/Cas9
multiplex genome editing
SlEF1α promoter
tomato
tRNA
資料タイプ
学術雑誌論文
抄録
Several expression systems for multiple guide RNA (gRNAs) have been developed in the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9) system to induce multiple-gene modifications in plants. Here, we evaluated mutation efficiencies in the tomato genome using multiplex CRISPR/Cas9 vectors consisting of various Cas9 expression promoters with multiple gRNA expression combinations. In transgenic tomato calli induced with these vectors, mutation patterns varied depending on the promoters used to express Cas9. By using the tomato ELONGATION FACTOR-1α (SlEF1α) promoter to drive Cas9, occurrence of various types of mutations with high efficiency was detected in the tomato genome. Furthermore, sequence analysis showed that the majority of mutations using the multiplex system with the SlEF1α promoter corresponded to specific mutation pattern of deletions produced by self-ligation at two target sites of CRISPR/Cas9 with low mosaic mutations. These results suggest that optimizing the Cas9 expression promoter used in CRISPR/Cas9-mediated mutation improves multiplex genome editing, and could be used effectively to disrupt functional domains precisely in the tomato genome.
掲載誌名
Frontiers in Plant Science
ISSN
1664462X
出版者
Frontiers Media S.A.
9
開始ページ
916
発行日
2018-07-03
権利情報
Copyright © 2018 Hashimoto, Ueta, Abe, Osakabe and Osakabe. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). (https://creativecommons.org/licenses/by/4.0/) The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
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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