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Microbial natural products (NPs) especially of the  Streptomyces  genus have been regarded as an unparalleled resource for pharmaceutical drugs discovery. Moreover, recent progress in sequencing technologies and computational resources further reinforces to identify numerous NP biosynthetic gene clusters (BGCs) from the genomes of  Streptomyces . However, the majority of these BGCs are silent or poorly expressed in native strains and remain to be activated and investigated, which relies heavily on efficient genome editing approaches. Accordingly, numerous strategies are developed, especially, the most recently developed, namely, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated (Cas) system reveals remarkable higher accuracy and efficiency for genome editing in various model organisms including the  Streptomyces . In this mini review, we highlight the application of CRISPR/Cas9-based approaches in  Streptomyces , focus on the editing of BGCs either  in vivo  or  in vitro , as well as target cloning of large-sized BGCs and heterologous expression in a genetically manipulatable host, for discovery, characterization, reengineering, and production of potential pharmaceutical drugs.

CRISPR/Cas9-Based Editing of Streptomyces for Discovery, Characterization, and Production of Natural Products