Journal
HORTICULTURE RESEARCH
Volume 9, Issue -, Pages -Publisher
OXFORD UNIV PRESS INC
DOI: 10.1093/hr/uhab004
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Funding
- Cabinet Office, Government of Japan, Cross-ministerial Strategic Innovation Promotion Program (SIP), Technologies for creating nextgeneration agriculture, forestry and fisheries (Bio-oriented Technology Research Advancement Institution, NARO)
- JSPS [18 K19200]
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Target-AID is a CRISPR/Cas9-based genome-editing tool that confers base-editing capability on the Cas9 system. In this study, we successfully applied the Target-AID system to edit negative regulators of the plant hormone signaling pathway in tomatoes, demonstrating its potential for molecular breeding in tomato crops.
Target activation-induced cytidine deaminase (Target-AID), a novel CRISPR/Cas9-based genome-editing tool, confers the base-editing capability on the Cas9 genome-editing system. It involves the fusion of cytidine deaminase (CDA), which catalyzes cytidine (C) to uridine (U) substitutions, to the mutated nickase-type nCas9 or deactivated-type dCas9. To confirm and extend the applicability of the Target-AID genome-editing system in tomatoes (Solanum lycopersicum L.), we transformed the model tomato cultivar Micro-Tom and commercial tomato cultivars using this system by targeting SlDELLA, which encodes a negative regulator of the plant phytohormone gibberellic acid (GA) signaling pathway. We confirmed that the nucleotide substitutions were induced by the Target-AID system, and we isolated mutants showing high GA sensitivity in both Micro-Tom and the commercial cultivars. Moreover, by successfully applying this system to ETHYLENE RECEPTOR 1 (SlETR1) with single sgRNA targeting, double sgRNA targeting, as well as dual-targeting of both SlETR1 and SlETR2 with a single sgRNA, we demonstrated that the Target-AID genome-editing system is a promising tool for molecular breeding in tomato crops. This study highlights an important aspect of the scientific and agricultural potential of the combinatorial use of the Target-AID and other base-editing systems.
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