articleACS Synthetic BiologyDec 2, 2014HYBRID OA

High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System

RERyan E. CobbYWYajie WangHZHuimin Zhao

University of Illinois Urbana-Champaign

PubMed
Indexed incrossrefpubmed

Abstract

Actinobacteria, particularly those of genus Streptomyces, remain invaluable hosts for the discovery and engineering of natural products and their cognate biosynthetic pathways. However, genetic manipulation of these bacteria is often labor and time intensive. Here, we present an engineered CRISPR/Cas system for rapid multiplex genome editing of Streptomyces strains, demonstrating targeted chromosomal deletions in three different Streptomyces species and of various sizes (ranging from 20 bp to 30 kb) with efficiency ranging from 70 to 100%. The designed pCRISPomyces plasmids are amenable to assembly of spacers and editing templates via Golden Gate assembly and isothermal assembly (or traditional…

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Topics & keywords

Topics
Keywords
  • Streptomyces
  • Biology
  • Genome editing
  • CRISPR
  • Plasmid
  • Genome
  • Genetics
  • Computational biology
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