Authors
Tien Van Vu, Gah-Hyun Lim, Seung Hee Choi, Ju Yeon Moon, Ngan Thi Nguyen, Swati Das, Mil Thi Tran, Yeon Woo Sung, Jihae Kim, Young Jong Song, Suk Weon Kim, Jae Cheol Jeong, Jae-Yean Kim
Publication date
2022/8/27
Journal
bioRxiv
Pages
2022.08. 27.505510
Publisher
Cold Spring Harbor Laboratory
Description
Highly efficient precision gene editing is ultimately a desirable technology, but implementation remains challenging, especially in plants. Here, we tested a novel CRISPR-Cas-based tool for precision gene replacement in plants employing a microhomology-mediated end joining (MMEJ) mechanism. The MMEJ-mediated precise gene replacement employing 20-bp microhomology MMEJ donors (~100 bp lengths) and a cNHEJ inhibitor, NU7441, produced high targeted editing efficiencies, up to 1.60 ± 0.14, 4.47 ± 1.98, and 8.98 ± 4.73 % in tomato, lettuce, and cabbage, respectively. Our data also revealed the critical impacts of the microhomology length and NU7441 concentration on MMEJ-based precision gene editing in plants.
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