Modification of tomato breeding traits and plant hormone signaling by Target-AID, the genome-editing system inducing efficient nucleotide substitution

Recently, scientists from the University of Tsukuba developed a novel CRISPR/Cas9-based genome-editing tool called Target activation-induced cytidine deaminase (Target-AID). They targeted SlDELLA and SlETR in the model tomato cultivar ‘Micro-Tom’ and in commercial tomato cultivars to confirm and extend the applicability of the Target-AID genome-editing system. After confirming that nucleotide substitutions were induced by the Target-AID system, mutants that showed high GA sensitivity in both ‘Micro-Tom’ and the commercial cultivars were isolated. Nucleotide substitution was also successfully induced by multi-targeting with a single sgRNA and multi-targeting with multiple sgRNAs in a single vector. In addition, the Target-AID system also produced a Target-AID-specific pattern of indels with editing positions different from those of Cas9 vectors.

“We demonstrate that the Target-AID system could efficiently edit sgRNA-targeting sites and facilitate a case study on the application of base-editing technology to translational research that aims at the commercialization of genome-edited crops”, Prof Ezura said. These results demonstrated that the Target-AID genome-editing system is a promising tool for molecular breeding in tomato crops, highlighting the scientific and agricultural potential of the combined use of Target-AID with other base-editing systems.

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Reference

Authors

Sachiko Kashojiya1,2,†, Yu Lu1,2,†, Mariko Takayama1, Hiroki Komatsu1, Luyen Hieu Thi Minh1, Keiji Nishida3,4, Kenta Shirasawa5, Kenji Miura1, Satoko Nonaka1, Jun-ichiro Masuda1,6, Akihiko Kondo3,4, Hiroshi Ezura1,7,* and Tohru Ariizumi1,7,*

Affiliations

1 Faculty of Life and Environmental Sciences, University of Tsukuba, Gene Research Center, Tsukuba, Ibaraki 305-8572, Japan

2 Japan Society for Promotion of Science, 5-3-1, Kojimachi, Tokyo 102-0083, Japan

3 Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, Hyogo 657-8501, Japan

4 Engineering Biology Research Center, Kobe University, 7-1-49, Minatojima Minami Machi, Chuo-ku, Kobe 650-0047, Japan

5 Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, Japan

6 Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki, Miyazaki 889-2192, Japan

7 Tsukuba Plant Innovation Research Center, University of Tsukuba, Ibaraki 305-8572, Japan

About Dr. Hiroshi Ezura and Dr. Tohru Ariizumi

Dr. Hiroshi Ezura is a professor in the Faculty of Life and Environmental Sciences at the University of Tsukuba in Japan, as well as the director of the Plant Innovation Research Center at the University of Tsukuba. Prof. Ezura received his Ph.D. from Hokkaido University in 1993 and has been working at the University of Tsukuba since 2005. Prof. Ezura is a member of the Japanese Science Committee and has served as the chairman of the Japanese Society for Plant Cell and Molecular Biology. He is mainly engaged in research on the functional genomics and genetic breeding of Solanaceae and Cucurbitaceae vegetable crops. His group used gene-editing technology to breed the first Japanese tomato variety with blood pressure–lowering effects. Prof. Ezura has published numerous papers in Nature Biotechnology, Plant Cell, New Phytologist, Plant Physiology, Plant Journal, and other journals.

Dr. Tohru Ariizumi graduated from the Faculty of Agriculture, Tohoku University, completed the applied life science course at the Graduate School of Agriculture, University of Tokyo in 2004, and became a Doctor of Agriculture (Tohoku University). He was an assistant professor at Washington State University and became an assistant professor at the Graduate School of Life and Environmental Sciences at Tsukuba University in 2015.