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Gene editing

CRISPR/Cas9

The bacteria and archaea adaptive immune systems clustered regularly interspaced short palindromic repeats (CRISPR)/Cas have been modified and engineered to efficiently, conveniently and precisely edit genomes in various cells and species both in and ex vivo. The gene editing toolkit is composed of two elements: the CRISPR associated protein (Cas9) and the guide RNA (gRNA). The guide RNA together with the genome matching sequence locate Cas9 nuclease to the target site demarcated by conserved sequences called proto-spacer adjacent motifs (PAMs). Under physiological conditions in bacteria and archaea, the exogenous phage genomes or plasmids that contain the target DNA sequences are cleaved and subjected to degradation. While the engineered systems  create site-specific double-stranded DNA breaks, which are then repaired either by homologous recombination or by non-homologous end joining.
 
Targeted genome editing with RNA-guided Cas9. (Rewriting a genome. NEWS & VIEWS, Nature ) 
The CRISPR/Cas9 system has been demonstrated to target multiplex editing for genome engineering, including gene knock-out, chromosome deletion and translocation, gene or promoter knock-in, gene activation and repression, epigenome modification, and etc.
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References
1. Tetsushi Sakuma, Ayami Nishikawa, Satoshi Kume, Kazuaki Chayama & Takashi Yamamoto.Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. 
2. Bastian Minkenberg, Matthew Wheatley, Yinong Yang. CRISPR/Cas9-Enabled Multiplex Genome Editing and Its Application. Progress in Molecular Biology and Translational Science, Volume 149 ISSN 1877-1173.
3. Le Cong, F. Ann Ran, David Cox, Shuailiang Lin, Robert Barretto,& Naomi Habib, Patrick D. Hsu, Xuebing Wu, Wenyan Jiang, Luciano A. Marraffini, Feng Zhang. Multiplex Genome Engineering Using CRISPR/Cas Systems. Science.
4. Zehua Bao, Han Xiao, Jing Liang, Lu Zhang, Xiong Xiong, Ning Sun, Tong Si, and Huimin Zhao. Homology-Integrated CRISPR-Cas (HI-CRISPR) System for One-Step Multigene Disruption in Saccharomyces cerevisia. dx.doi.org/10.1021/sb500255k | ACS Synth. Biol. 2015, 4, 585-594.  



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