Work

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

Year: 2016

Type: review

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.

Source: International Journal of Molecular Sciences

DOI: 10.3390/ijms17091507

Authors: Francisco Martin, Sabina Sánchez-Hernández, Alejandra Gutiérrez-Guerrero, Javier Pinedo-Gomez, Karim Benabdellah

Institutions: Universidad de Granada, Pfizer-University of Granada-Junta de Andalucía Centre for Genomics and Oncological Research

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $3,499