Tandem Paired Nicking Promotes Precise Genome Editing with Scarce Interference by p53

• DOI: 10.1016/j.celrep.2019.12.064
• Publication Date: 2020-01-28T16:05:57Z
• Authors: Toshinori Hyodo; Md. Lutfur Rahman; Sivasundaram Karnan; Takuji Ito; Atsushi Toyoda; Akinobu Ota; Md Wahiduzzaman; Shinobu Tsuzuki; Yohei Okada; Yoshitaka Hosokawa; Hiroyuki Konishi
• Keywords: Tandem

Highlights•Tandem paired nicking promotes targeted knockin with substantial efficiency•Tandem scarcely creates indels at the edited genomic loci•DNA recombination elicited by tandem is analogous to Holliday's model•Targeted shows no reciprocal interference p53SummaryTargeted mediated double-stranded DNA cleavage accompanied unwanted insertions and deletions (indels) on-target off-target sites. A nick-mediated approach generates but exhibits reduced efficiency of knockin. Here, we demonstrate that nicking, a method for involving two Cas9 nickases create nicks homologous regions donor genome in same strand, loci, while permitting largely equivalent Cas9-nuclease-based approach. Tandem seems accomplish model intended changes without introducing additional nucleotide changes, such as silent mutations. Targeted through neither triggers significant p53 activation nor occurs preferentially p53-suppressed cells. These properties its utility precision engineering.Graphical abstract