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Int. J. Mol. Sci. 2016, 17(9), 1507; doi:10.3390/ijms17091507

Biased and Unbiased Methods for the Detection of Off-Target Cleavage by CRISPR/Cas9: An Overview

1
Genomic Medicine Department, GENYO—Centre for Genomics and Oncological Research Pfizer-Universidad de Granada-Junta de Andalucía, Avda de la Ilustración 114, 18007 Granada, Spain
2
LentiStem Biotech, GENYO, Avda de la Ilustración 114, 18007 Granada, Spain
*
Authors to whom correspondence should be addressed.
Academic Editor: Izuho Hatada
Received: 29 June 2016 / Revised: 29 August 2016 / Accepted: 31 August 2016 / Published: 8 September 2016
(This article belongs to the Special Issue Genome Editing)
View Full-Text   |   Download PDF [982 KB, uploaded 8 September 2016]   |  

Abstract

The clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein 9 endonuclease (Cas9) derived from bacterial adaptive immune systems is a revolutionary tool used in both basic and applied science. It is a versatile system that enables the genome of different species to be modified by generating double strand breaks (DSBs) at specific locations. However, all of the CRISPR/Cas9 systems can also produce DSBs at off-target sites that differ substantially from on-target sites. The generation of DSBs in locations outside the intended site can produce mutations that need to be carefully monitored, especially when using these tools for therapeutic purposes. However, off-target analyses of the CRISPR/Cas9 system have been very challenging, particularly when performed directly in cells. In this manuscript, we review the different strategies developed to identify off-targets generated by CRISPR/cas9 systems and other specific nucleases (ZFNs, TALENs) in real target cells. View Full-Text
Keywords: specific nucleases; CRISPR/cas9; off-target sites; real target cells; unbiased; biased; CHIPSeq; IDLVs; GUIDE-seq; LAM-HTGTS specific nucleases; CRISPR/cas9; off-target sites; real target cells; unbiased; biased; CHIPSeq; IDLVs; GUIDE-seq; LAM-HTGTS
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Martin, F.; Sánchez-Hernández, S.; Gutiérrez-Guerrero, A.; Pinedo-Gomez, J.; Benabdellah, K. Biased and Unbiased Methods for the Detection of Off-Target Cleavage by CRISPR/Cas9: An Overview. Int. J. Mol. Sci. 2016, 17, 1507.

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