Journal
MOLECULAR THERAPY-NUCLEIC ACIDS
Volume 4, Issue -, Pages -Publisher
CELL PRESS
DOI: 10.1038/mtna.2015.37
Keywords
CRISPR/Cas9; off-target
Categories
Funding
- Ministry of Science of China [2012CBA01302]
- Science and Technology Department of Guangdong Province, China [2014B020225007]
- Program for New Century Excellent Talents in University [NCET-121078]
- National Natural Science Foundation of China [31071279]
- Science and Technology Innovation Project Grant from the Department of Education of Guangdong Province [2013KJCX0029]
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CRISPR/Cas9 is a versatile genome-editing technology that is widely used for studying the functionality of genetic elements, creating genetically modified organisms as well as preclinical research of genetic disorders. However, the high frequency of off-target activity (>= 50%)-RGEN (RNA-guided endonuclease)-induced mutations at sites other than the intended on-target site-is one major concern, especially for therapeutic and clinical applications. Here, we review the basic mechanisms underlying off-target cutting in the CRISPR/Cas9 system, methods for detecting off-target mutations, and strategies for minimizing off-target cleavage. The improvement off-target specificity in the CRISPR/Cas9 system will provide solid genotype-phenotype correlations, and thus enable faithful interpretation of genome-editing data, which will certainly facilitate the basic and clinical application of this technology.
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