Nicking Endonuclease-Mediated Vector Construction Strategies for Plant Gene Functional Research.

ABSTRACT: Plant genetic engineering vectors, such as RNA interference (RNAi) and CRISPR/Cas9 vectors, are important tools for plant functional genomics. Efficient construction of these functional vectors can facilitate the study of gene function. Although some methods for vector construction have been reported, their operations are still complicated and costly. Here, we describe a simpler and low-cost vector construction method by nicking endonucleases-mediated DNA assembly (NEMDA), which uses nicking endonucleases to generate single-strand overhanging complementary ends for rapid assembly of DNA fragments into plasmids. Using this approach, we rapidly completed the construction of four RNAi vectors and a CRISPR/Cas9 knockout vector with five single-guide RNA (sgRNA)-expression cassettes for multipl

SUBMITTER: Gong Q 

PROVIDER: S-EPMC7570347 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature