Johan Hunziker, Keiji Nishida, Akihiko Kondo, Sanae Kishimoto, Tohru Ariizumi & Hiroshi Ezura

Scientific Reports volume 10, Article number: 20471 (2020) 

The use of Target activation-induced cytidine deaminase (Target-AID) base-editing technology with the CRISPR-Cas 9 system fused with activation-induced cytidine deaminase (AID) resulted in the substitution of a cytidine with a thymine. In previous experiments focusing on a single target gene, this system has been reported to work in several plant species, including tomato (Solanum lycopersicum L.). In this research, we used Target-AID technology to target multiple genes related to carotenoid accumulation in tomato. We selected 3 genes, SlDDB1, SlDET1 and SlCYC-B, for their roles in carotenoid accumulation. Among 12 edited T₀ lines, we obtained 10 independent T₀ lines carrying nucleotide substitutions in the three targeted genes, with several allelic versions for each targeted gene. The two edited lines showed significant differences in carotenoid accumulation. These results demonstrate that Target-AID technology is a highly efficient tool for targeting multiple genes with several allelic versions.

See https://www.nature.com/articles/s41598-020-77379-2

Figure 1: Representation of SlDDB1, SlDET1 and SlCYC-B target sites. Schematic representation of each candidate gene, (a) SlDDB1, (b) SlDET1 and (c) SlCYC-B, with the previously described mutations and their corresponding annotations with mutation position compared to the first nucleotide of start codon on gene sequence. Black boxes represent exons regions and lines represent introns or UTR regions. Double slashes in introns represent reduction of the intron size. The red triangles represent the site targeted by the Target-AID with their corresponding nucleotides localization on gene.