OF, off-target. Gene names, amino acid mutations and single nucleotide conversions are indicated by blue, red and green, respectively. c, Sanger sequencing chromatograms show that U to C mutation was observed only in the RNA of three ABE7.10 off-target sites. Extended Data Fig. 5 Biotypes...
Recently developed DNA base editing methods enable the direct generation of desired point mutations in genomic DNA without generating any double-strand breaks1–3, but the issue of off-target edits has limited the application of these methods. Although s
Off-target DNA mutationOff-target RNA mutationBase editors (BEs) is a powerful tool that could edit the genome efficiently without inducing DNA double-strand breaks (DSBs). Recently, off-target DNA and RNA mutations of BEs were found, rasing concerns about its precision and safety. Here we ...
Off-target RNA mutation induced by DNA base editing and its elimination by mutagenesis. Nature 571, 275–278 (2019). Article Google Scholar Rees, H. A., Wilson, C., Doman, J. L. & Liu, D. R. Analysis and minimization of cellular RNA editing by DNA adenine base editors. Sci. Adv...
To assess the off-target mutation rate in grapevine (Vitis vinifera), we performed whole-genome sequencing (WGS) of seven Cas9-edited grapevine plants in which one of two genes was targeted by CRISPR/Cas9 and three wild-type (WT) plants. In total, we identified between 202,008 and 272,...
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...
Although RNA-guided genome editing via the CRISPR-Cas9 system is now widely used in biomedical research, genome-wide target specificities of Cas9 nucleases remain controversial. Here we present Digenome-seq, in vitro Cas9-digested whole-genome sequencing, to profile genome-wide Cas9 off-target effect...
CRISPR/ Cas9 technique, as the third generation of gene editing tool, has been widely used for genome editing in varous cells and organisms. Despite the efficient of gene mutation triggered by guide RNA/ cas9, the specificity...
identifying most off-targets with mutation rates superior to 0.1 %, while the number of false positives can be largely reduced with a cutoff on the off-target score. We also evaluate on-target efficiency prediction algorithms against available datasets. The correlation between the predictions and th...
et al. Off-target RNA mutation induced by DNA base editing and its elimination by mutagenesis. Nature 571, 275–278 (2019). 19. Cameron, P. et al. Mapping the genomic landscape of CRISPR–Cas9 cleavage. Nat. Methods 14, 600–606 (2017...