Modification of tomato breeding traits and plant hormone signaling by Target-AID, the genome-editing system inducing efficient nucleotide substitution. Issue 1 (28th January 2022)

• Record Type: Journal Article
• Authors: Kashojiya, Sachiko; Lu, Yu; Takayama, Mariko; Komatsu, Hiroki; Minh, Luyen Hieu Thi; Nishida, Keiji; Shirasawa, Kenta; Miura, Kenji; Nonaka, Satoko; Masuda, Jun-ichiro; Kondo, Akihiko; Ezura, Hiroshi; Ariizumi, Tohru
• Is Part Of: Horticulture research. Volume 9:Issue 1(2022)
• Publication Date: 2022-01-28
• Subjects: Horticulture -- Research -- Periodicals; 635.072
• DOI: 10.1093/hr/uhab004
• Languages: English
• ISSNs: 2052-7276

Abstract

Abstract: 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.