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Curr Opin Microbiol. 2014 Jun;19:114-119. doi: 10.1016/j.mib.2014.07.001. Epub 2014 Jul 19.

Harnessing CRISPR-Cas9 immunity for genetic engineering.

Author information

1
Helmholtz Centre for Infection Research, Department of Regulation in Infection Biology, Braunschweig 38124, Germany; The Laboratory for Molecular Infection Medicine Sweden (MIMS), Umeå Centre for Microbial Research (UCMR), Department of Molecular Biology, Umeå University, Umeå 90187, Sweden; Hannover Medical School, Hannover 30625, Germany. Electronic address: emmanuelle.charpentier@helmholtz-hzi.de.
2
Laboratory of Bacteriology, The Rockefeller University, 1230 York Ave., New York, NY 10065, USA. Electronic address: marraffini@rockefeller.edu.

Abstract

CRISPR-Cas encodes an adaptive immune system that defends prokaryotes against infectious viruses and plasmids. Immunity is mediated by Cas nucleases, which use small RNA guides (the crRNAs) to specify a cleavage site within the genome of invading nucleic acids. In type II CRISPR-Cas systems, the DNA-cleaving activity is performed by a single enzyme Cas9 guided by an RNA duplex. Using synthetic single RNA guides, Cas9 can be reprogrammed to create specific double-stranded DNA breaks in the genomes of a variety of organisms, ranging from human cells to bacteria, and thus constitutes a powerful tool for genetic engineering. Here we describe recent advancements in our understanding of type II CRISPR-Cas immunity and how these studies led to revolutionary genome editing applications.

PMID:
25048165
PMCID:
PMC4155128
DOI:
10.1016/j.mib.2014.07.001
[Indexed for MEDLINE]
Free PMC Article

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