Genome editing in plants using CRISPR type I-D nuclease

Communications Biology - Tập 3 Số 1
Keishi Osakabe1, Naoki Wada1, Tomoko Miyaji1, Emi Murakami1, Kazuya Marui1, Risa Ueta1, Ryosuke Hashimoto1, Chihiro Abe-Hara1, Bihe Kong2, Kentaro Yano2, Yuriko Osakabe1
1Graduate School of Technology, Industrial and Social Sciences, Tokushima University, Tokushima, 770-8503, Japan
2Department of Life Sciences, School of Agriculture, Meiji University, Kanagawa, 214-8571, Japan

Tóm tắt

AbstractGenome editing in plants has advanced greatly by applying the clustered regularly interspaced short palindromic repeats (CRISPRs)-Cas system, especially CRISPR-Cas9. However, CRISPR type I—the most abundant CRISPR system in bacteria—has not been exploited for plant genome modification. In type I CRISPR-Cas systems, e.g., type I-E, Cas3 nucleases degrade the target DNA in mammals. Here, we present a type I-D (TiD) CRISPR-Cas genome editing system in plants. TiD lacks the Cas3 nuclease domain; instead, Cas10d is the functional nuclease in vivo. TiD was active in targeted mutagenesis of tomato genomic DNA. The mutations generated by TiD differed from those of CRISPR/Cas9; both bi-directional long-range deletions and short indels mutations were detected in tomato cells. Furthermore, TiD can be used to efficiently generate bi-allelic mutant plants in the first generation. These findings indicate that TiD is a unique CRISPR system that can be used for genome engineering in plants.

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Communications Biology

Tập 3 Số 1

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