Format

Send to

Choose Destination
Methods Enzymol. 2016;575:271-84. doi: 10.1016/bs.mie.2016.03.014. Epub 2016 Mar 29.

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

Author information

1
University of Illinois at Urbana-Champaign, Urbana, IL, United States.
2
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.
3
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.

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.

KEYWORDS:

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

PMID:
27417933
DOI:
10.1016/bs.mie.2016.03.014
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center