Coevolutionary Couplings Unravel PAM-Proximal Constraints of CRISPR-SpCas9.

ABSTRACT: The clustered regularly interspaced short palindromic repeats (CRISPR) system, an immune system analog found in prokaryotes, allows a single-guide RNA to direct a CRISPR-associated protein (Cas) with combined helicase and nuclease activity to DNA. The presence of a specific protospacer adjacent motif (PAM) next to the DNA target site plays a crucial role in determining both efficacy and specificity of gene editing. Herein, we introduce a coevolutionary framework to computationally unveil nonobvious molecular interactions in CRISPR systems and experimentally probe their functional role. Specifically, we use direct coupling analysis, a statistical inference framework used to infer direct coevolutionary couplings, in the context of protein/nucleic acid interactions. Applied to Streptococcus p

SUBMITTER: Li Y 

PROVIDER: S-EPMC6838953 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature