In-situ generation of large numbers of genetic combinations for metabolic reprogramming via CRISPR-guided base editing.

ABSTRACT: Reprogramming complex cellular metabolism requires simultaneous regulation of multigene expression. Ex-situ cloning-based methods are commonly used, but the target gene number and combinatorial library size are severely limited by cloning and transformation efficiencies. In-situ methods such as multiplex automated genome engineering (MAGE) depends on high-efficiency transformation and incorporation of heterologous DNA donors, which are limited to few microorganisms. Here, we describe a Base Editor-Targeted and Template-free Expression Regulation (BETTER) method for simultaneously diversifying multigene expression. BETTER repurposes CRISPR-guided base editors and in-situ generates large numbers of genetic combinations of diverse ribosome binding sites, 5' untranslated regions, or promoters,

SUBMITTER: Wang Y 

PROVIDER: S-EPMC7846839 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature