Expanding the Scope of Bacterial CRISPR Activation with PAM-Flexible dCas9 Variants.

ABSTRACT: CRISPR-Cas transcriptional tools have been widely applied for programmable regulation of complex biological networks. In comparison to eukaryotic systems, bacterial CRISPR activation (CRISPRa) has stringent target site requirements for effective gene activation. While genes may not always have an NGG protospacer adjacent motif (PAM) at the appropriate position, PAM-flexible dCas9 variants can expand the range of targetable sites. Here we systematically evaluate a panel of PAM-flexible dCas9 variants for their ability to activate bacterial genes. We observe that dxCas9-NG provides a high dynamic range of gene activation for sites with NGN PAMs while dSpRY permits modest activity across almost any PAM. Similar trends were observed for heterologous and endogenous promoters. For all variants t

SUBMITTER: Kiattisewee C 

PROVIDER: S-EPMC10516241 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature