Anthocyanin-assisted <italic>Agrobacterium</italic> infiltration for the rapid evaluation of genome editing efficiencies across multiple plant species

Abstract

CRISPR-based genome editing technologies continue to drive major advances in life sciences. A major challenge for realizing widespread use of genome editing in plants and agriculture is establishing methods that enable the rapid, comprehensive, and precise evaluation of editing technologies using transient methods. Here we report a new and rapid genome editing evaluation method using Agrobacterium infiltration techniques to enable broad-spectrum, simplistic, and precise assessments of genome editing efficiencies. We employed an anthocyanin marker to facilitate visual screenings of genome-edited cells for use in adult strawberry fruits as well as tomato fruits, cotton leaves, and sugar beet leaves. Using this method, we demonstrate the ability to quickly measure genome editing efficiencies mediated by SpCas9, LbCas12a, A3A-PBE, ABE8e, and PPE. This new method will allow researchers to rapidly and easily evaluate genome editing tools across a broad spectrum of plant species, further expediting the development of genome-edited agricultural crops.

Keywords

• genome editing
• <italic>Agrobacterium</italic> infiltration
• anthocyanin
• transient transformation
• geminivirus replicon
• strawberry
• tomato