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Annu Rev Biochem. 2016 Jun 2;85:227-64. doi: 10.1146/annurev-biochem-060815-014607. Epub 2016 Apr 25.

CRISPR/Cas9 in Genome Editing and Beyond.

Wang H1, La Russa M1,2, Qi LS1,3,4.

Author information

1
Department of Bioengineering, Stanford University, Stanford, California 94305; email: hfgwang@stanford.edu , mlarussa@stanford.edu , stanley.qi@stanford.edu.
2
Biomedical Sciences Graduate Program, University of California, San Francisco, California 94158.
3
Department of Chemical and Systems Biology, Stanford University, Stanford, California 94305.
4
Chemistry, Engineering and Medicine for Human Health (ChEM-H), Stanford University, Stanford, California 94305.

Abstract

The Cas9 protein (CRISPR-associated protein 9), derived from type II CRISPR (clustered regularly interspaced short palindromic repeats) bacterial immune systems, is emerging as a powerful tool for engineering the genome in diverse organisms. As an RNA-guided DNA endonuclease, Cas9 can be easily programmed to target new sites by altering its guide RNA sequence, and its development as a tool has made sequence-specific gene editing several magnitudes easier. The nuclease-deactivated form of Cas9 further provides a versatile RNA-guided DNA-targeting platform for regulating and imaging the genome, as well as for rewriting the epigenetic status, all in a sequence-specific manner. With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics. In this review, we describe the current models of Cas9 function and the structural and biochemical studies that support it. We focus on the applications of Cas9 for genome editing, regulation, and imaging, discuss other possible applications and some technical considerations, and highlight the many advantages that CRISPR/Cas9 technology offers.

KEYWORDS:

CRISPR applications; Cas9 structure; dCas9; epigenetic regulation; gene regulation; genomic imaging

[Indexed for MEDLINE]

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