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

被引:57
|
作者
Martin, Francisco [1 ,2 ]
Sanchez-Hernandez, Sabina [1 ]
Gutierrez-Guerrero, Alejandra [1 ]
Pinedo-Gomez, Javier [1 ]
Benabdellah, Karim [1 ,2 ]
机构
[1] Pfizer Univ Granada Junta de Andalucia, Genom Med Dept, GENYO Ctr Genom & Oncol Res, Avda Ilustrac 114, Granada 18007, Spain
[2] GENYO, LentiStem Biotech, Avda Ilustrac 114, Granada 18007, Spain
来源
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2016年 / 17卷 / 09期
关键词
specific nucleases; CRISPR; cas9; off-target sites; real target cells; unbiased; biased; CHIPSeq; IDLVs; GUIDE-seq; LAM-HTGTS; GENOME-WIDE ANALYSIS; CHIP-SEQ; CRISPR-CAS9; CAS9; REPEATS; SITES; MUTATIONS; NUCLEASES; TALENS; GENES;
D O I
10.3390/ijms17091507
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
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页数:9
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