Rapid, modular, and cost-effective generation of donor DNA constructs for CRISPR-based gene knock-in.

ABSTRACT: Clustered regularly interspaced short palindromic repeats (CRISPR)-based gene editing techniques find applications in many fields, such as molecular biology, cancer biology, and disease modeling. In contrast to the knock-out procedure, a key step of CRISPR knock-in experiments is the homology-directed repair process that requires donor constructs as repair templates. Therefore, it is desirable to generate a series of donor templates efficiently and cost-effectively. In this study, we developed a new strategy that combines (i) Gibson assembly reaction, (ii) a linker pair composed of eight in silico screened restriction enzyme sites, and (iii) a hierarchical framework, to remarkably improve the efficiency of producing donor constructs for common genes as well as for the genes containing unba

SUBMITTER: Chen YJ 

PROVIDER: S-EPMC7211398 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature