Osakabe, Keishi Tokushima University Tokushima University Educator and Researcher Directory KAKEN Search Researchers
Wada, Naoki Tokushima University Tokushima University Educator and Researcher Directory
Miyaji, Tomoko Tokushima University
Murakami, Emi Tokushima University
Marui, Kazuya Tokushima University
Ueta, Risa Tokushima University
Hashimoto, Ryosuke Tokushima University
Abe-Hara, Chihiro Tokushima University
Kong, Bihe Meiji University
Yano, Kentaro Meiji University
Genome 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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cb_3_648.pdf 3.05 MB
Bioscience and Bioindustry