Progress of application and off-target effects of CRISPR/Cas9.
The clustered regulatory interspaced short palindromic repeat/Cas9 (CRISPR/Cas9) system mediates genome editing and is revolutionizing genetic researches. Scientists are able to manipulate the gene of interest from any organism with CRISPR/Cas9. Compared with zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) technologies, the CRISPR/Cas9 technology provides an easy and efficient approach to manipulate the genome. In this system, sgRNA (Single guide RNA), a short RNA matching the targeted DNA fragment, guides the CRISPR/Cas9 to interrogate the genome. Because sgRNA can tolerate certain mismatches to the DNA targets and thereby promote undesired off-target mutagenesis, the key limit of this technology is off-target effects. To eliminate the off-target effects, different strategies have been adopted. In this review, we summarize the application of CRISPR/Cas9 and different strategies for addressing off-target effects.
State Key Laboratory Cultivating Base of Optometry and Visual Science, Key Laboratory of Vision Science, Ministry of Health, The Eye Hospital of Wenzhou Medical University, Wenzhou 325027 China.
Wu
W
Zheng
State Key Laboratory Cultivating Base of Optometry and Visual Science, Key Laboratory of Vision Science, Ministry of Health, The Eye Hospital of Wenzhou Medical University, Wenzhou 325027 China.
Feng
F
Gu
D000818:Animals; D035843:Biomedical Research / Q000379:methods / Q000639:trends; D064113:CRISPR-Cas Systems; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D005544:Forecasting; D018390:Gene Targeting / Q000379:methods* / Q000639:trends; D005818:Genetic Engineering / Q000379:methods* / Q000639:trends; D006801:Humans; D008957:Models, Genetic; D015203:Reproducibility of Results
D004247:DNA