title: High-Efficiency Genome Editing of Streptomyces Species by an Engineered CRISPR/Cas System.
abstract: Next-generation sequencing technologies have rapidly expanded the genomic information of numerous organisms and revealed a rich reservoir of natural product gene clusters from microbial genomes, especially from Streptomyces, the largest genus of known actinobacteria at present. However, genetic engineering of these bacteria is often time consuming and labor intensive, if even possible. In this chapter, we describe the design and construction of pCRISPomyces, an engineered Type II CRISPR/Cas system, for targeted multiplex gene deletions in Streptomyces lividans, Streptomyces albus, and Streptomyces viridochromogenes with editing efficiency ranging from 70% to 100%. We demonstrate pCRISPomyces as a powerful tool for genome editing in Streptomyces.
authors.affiliation: University of Illinois at Urbana-Champaign, Urbana, IL, United States.
authors.forename: Y
authors.initials: Y
authors.lastname: Wang
authors.affiliation: University of Illinois at Urbana-Champaign, Urbana, IL, United States; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, United States.
authors.forename: R E
authors.initials: RE
authors.lastname: Cobb
authors.affiliation: University of Illinois at Urbana-Champaign, Urbana, IL, United States; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, United States. Electronic address: zhao5@illinois.edu.
authors.forename: H
authors.initials: H
authors.lastname: Zhao
mesh_terms: D001688:Biological Products / Q000378:metabolism; D053898:Biosynthetic Pathways; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D017353:Gene Deletion; D000072669:Gene Editing / Q000379:methods*; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
chemical_list: D001688:Biological Products
keywords: CRISPR; Genome engineering; Natural product biosynthesis; Streptomyces; Synthetic biology