CRISPR/Cas-based precision gene replacement in plants via microhomology-mediated end joining

Autor: 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
Rok vydání: 2022
Popis: 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.
Databáze: OpenAIRE