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

Y. Wang, R. E. Cobb, Huimin Zhao

Research output: Chapter in Book/Report/Conference proceedingChapter

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.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
EditorsSarah E. O'Connor
PublisherAcademic Press Inc.
Pages271-284
Number of pages14
DOIs
StatePublished - 2016

Publication series

NameMethods in Enzymology
Volume575
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Fingerprint

CRISPR-Cas Systems
Clustered Regularly Interspaced Short Palindromic Repeats
Streptomyces
Genes
Microbial Genome
Streptomyces lividans
Genetic engineering
Genetic Engineering
Actinobacteria
Gene Deletion
Multigene Family
Biological Products
Bacteria
Personnel
Technology
Gene Editing

Keywords

  • CRISPR
  • Genome engineering
  • Natural product biosynthesis
  • Streptomyces
  • Synthetic biology

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Wang, Y., Cobb, R. E., & Zhao, H. (2016). High-Efficiency Genome Editing of Streptomyces Species by an Engineered CRISPR/Cas System. In S. E. O'Connor (Ed.), Methods in Enzymology (pp. 271-284). (Methods in Enzymology; Vol. 575). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2016.03.014

High-Efficiency Genome Editing of Streptomyces Species by an Engineered CRISPR/Cas System. / Wang, Y.; Cobb, R. E.; Zhao, Huimin.

Methods in Enzymology. ed. / Sarah E. O'Connor. Academic Press Inc., 2016. p. 271-284 (Methods in Enzymology; Vol. 575).

Research output: Chapter in Book/Report/Conference proceedingChapter

Wang, Y, Cobb, RE & Zhao, H 2016, High-Efficiency Genome Editing of Streptomyces Species by an Engineered CRISPR/Cas System. in SE O'Connor (ed.), Methods in Enzymology. Methods in Enzymology, vol. 575, Academic Press Inc., pp. 271-284. https://doi.org/10.1016/bs.mie.2016.03.014
Wang Y, Cobb RE, Zhao H. High-Efficiency Genome Editing of Streptomyces Species by an Engineered CRISPR/Cas System. In O'Connor SE, editor, Methods in Enzymology. Academic Press Inc. 2016. p. 271-284. (Methods in Enzymology). https://doi.org/10.1016/bs.mie.2016.03.014
Wang, Y. ; Cobb, R. E. ; Zhao, Huimin. / High-Efficiency Genome Editing of Streptomyces Species by an Engineered CRISPR/Cas System. Methods in Enzymology. editor / Sarah E. O'Connor. Academic Press Inc., 2016. pp. 271-284 (Methods in Enzymology).
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