Efficient, Precise Multiplex Genome Editing Developed in TomatoSeptember 12, 2018
Multiplex CRISPR-Cas9 has been used in some crops, but promoter optimization is hardly reported. Researcher Ryosuke Hashimoto and colleagues from Tokushima University in Japan used various Cas9 expression promoters with different gRNA expression combinations to edit genes in tomato.
The researchers design "all-in-one" plasmids with different promoters for multiplex gene editing and selected transformed calli through GFP fluorescence. They find varying promoter-dependent mutation patterns among the tomato calli employed with the designed plasmids via PCR. Among the designed promoters, the tomato ELONGATION FACTOR-1α (SlEF1α) promoter drove the highest efficiency for CRISPR-Cas9 genome editing, with specific mutation patterns produced. These results highlight promoter optimization for CRISPR-Cas9 editing will enable precise disruptions of functional domains in tomato.
For more information, read the article in Frontiers in Plant Science.
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