RECC: A Red/ET-CRISPR/Cas9-based system enabling genome mining of marine <i>Pseudoalteromonas</i> for novel natural products.

ABSTRACT: Marine microorganisms possess vast biosynthetic potential, yet most of their biosynthetic gene clusters (BGCs) remain transcriptionally silent under laboratory conditions. Genetic intractability has been a major barrier to activating these cryptic pathways. Here, we present RECC, an integrated Red/ET-CRISPR/Cas9 system that enables seamless, marker-free genome editing in marine bacteria. RECC couples Red/ET recombineering with CRISPR/Cas9-mediated cleavage, allowing the incorporation of homology arms and protospacers into a single construct through one-step Gibson assembly, thereby substantially simplifying the engineering process. Using Pseudoalteromonas flavipulchra DSM 14401 as a model, we employed RECC to replace the native promoter of a silent nonribosomal peptide synthetase-po

SUBMITTER: Wang ZJ 

PROVIDER: S-EPMC12819058 | biostudies-literature | 2026 Jun

REPOSITORIES: biostudies-literature