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Curr Opin Biotechnol. 2016 Feb;37:61-68. doi: 10.1016/j.copbio.2015.10.003. Epub 2015 Nov 26.

Exploiting CRISPR-Cas immune systems for genome editing in bacteria.

Author information

1
Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, Raleigh, NC 27695, USA. Electronic address: rbarran@ncsu.edu.
2
Department of Food Science, University of Wisconsin-Madison, Madison, WI 53706, USA. Electronic address: vanpijkeren@wisc.edu.

Abstract

The CRISPR-Cas immune system is a DNA-encoded, RNA-mediated, DNA-targeting defense mechanism, which provides sequence-specific targeting of DNA. This molecular machinery can be engineered into the sgRNA:Cas9 technology, for programmable cleavage of DNA. Following the genesis of double-stranded DNA breaks, the DNA repair machinery generates mutations at the cleavage site using various pathways. This technology has revolutionized eukaryotic genome editing, and we are at the cusp of full exploitation in bacteria. Here, we discuss the potential of CRISPR-based technologies for use in bacteria, and highlight the application of single stranded DNA recombineering combined with CRISPR-Cas selection to edit the genome of a probiotic organism. We envision that CRISPR-Cas technologies will play a key role in the development of next-generation industrial bacteria.

PMID:
26629846
DOI:
10.1016/j.copbio.2015.10.003
[Indexed for MEDLINE]

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