Low incidence of off-target mutations in individual CRISPR-Cas9 and TALEN targeted human stem cell clones detected by whole-genome sequencing. Cell Stem Cell. 2014; 15(1):27-30. [PubMed: 24996167]A. Veres, B.S. Gosis, Q. Ding, R. Collins, A. Ragavendran, H. Brand, S. Erdin, ...
WASHINGTON, Feb. 28 (Xinhua) -- Chinese scientists found that a popular gene-editing tool may lead to wide off-target mutations, warning of serious clinical safety risks of the gene-editing technique. The study published on Thursday in the journal Science showed that a type of base editor us...
Recently developed DNA base editing methods enable the direct generation of desired point mutations in genomic DNA without generating any double-strand breaks1,2,3, but the issue of off-target edits has limited the application of these methods. Although several previous studies have evaluated off-ta...
19, and there is considerable interest in understanding the factors that dictate the number and positions of off-target mutations13,19. At present, methods such as targeted sequencing, exome sequencing, WGS (whole genome sequencing), BLESS (direct in situ breaks labeling, enrichment on streptavidin...
Increasing genome-editing specificity entails developing improved design methods that better predict the prevalence of off-target mutations as a function of genome composition and design of the engineered ribonucleoprotein (RNP). Early results from CRISPR-Cas9 genome editing in plant systems indicate that...
A team of Chinese scientists has now conducted the first full assessment of the effects of CRISPR-Cas9 on non-human primates to ascertain whether it results in off-target mutations. A controversial study published last year suggested CRISPR could introduce unintended mutations, but after several ...
Interestingly, such off-target mutations were caused by the same two gRNAs either with TadCBEa or TadDE (Supplementary Fig. 21), suggesting gRNA-dependent off-target mutations by these base editors were simply caused by high sequence similarity between the target site and the off-target sites, ...
We validated off-target sites at which insertions or deletions were induced with frequencies below 0.1%, near the detection limit of targeted deep sequencing. We also showed that Cas9 nucleases can be highly specific, inducing off-target mutations at merely several, rather than thousands of, sites...
However, it is difficult to determine off-target effects of editing due to single-nucleotide polymorphism in individuals. Here we developed a method named GOTI (genome-wide off-target analysis by two-cell embryo injection) to detect off-target mutations by editing one blastomere of two-cell ...
In addition, paired Cas9 nickases, composed of D10A Cas9 and guide RNA, which generate two single-strand breaks (SSBs) or nicks on different DNA strands, were highly specific in human cells, avoiding off-target mutations without sacrificing genome-editing efficiency. Interestingly, paired nickase...