Prime editing is an adaptation of the CRISPR-Cas system that uses a Cas9(H840A)-reverse transcriptase fusion and a guide RNA amended with template and primer binding site sequences to achieve RNA-templated conversion of the target DNA, allowing specified substitutions, insertions, and deletions. In the first report of prime editing in plants, a variety of edits in rice and wheat were described, including insertions up to 15 bp. Several studies in rice quickly followed, but none reported a larger insertion. Here, we report easy-to-use vectors for prime editing in dicots as well as monocots, their validation in Nicotiana benthamiana, rice, and Arabidopsis, and an insertion of 66 bp that enabled split-GFP fluorescent tagging.
Keywords: Arabidopsis; Nicotiana benthamiana; Oryza sativa; fluorescent tagging; plant genome editing; prime editing; split GFP.
Copyright © 2021 Wang, Kaya, Zhang, Rai, Willmann, Carpenter, Read, Martin, Fei, Leach, Martin and Bogdanove.