RT Journal Article SR Electronic T1 Tandem paired nicking promotes precise genome editing with scarce interference by p53 JF bioRxiv FD Cold Spring Harbor Laboratory SP 810333 DO 10.1101/810333 A1 Toshinori Hyodo A1 Md Lutfur Rahman A1 Sivasundaram Karnan A1 Takuji Ito A1 Atsushi Toyoda A1 Akinobu Ota A1 Md Wahiduzzaman A1 Shinobu Tsuzuki A1 Yohei Okada A1 Yoshitaka Hosokawa A1 Hiroyuki Konishi YR 2019 UL http://biorxiv.org/content/early/2019/10/18/810333.abstract AB Targeted knock-in mediated by double-stranded DNA cleavage is accompanied by unwanted insertions and deletions (indels) at on-target and off-target sites. A nick-mediated approach scarcely generates indels but exhibits reduced efficiency of targeted knock-in. Here, we demonstrate that tandem paired nicking, a method for targeted knock-in involving two Cas9 nickases that create nicks at the homologous regions of the donor DNA and the genome in the same strand, scarcely creates indels at the edited genomic loci, while permitting the efficiency of targeted knock-in largely equivalent to that of the Cas9 nuclease-based approach. Tandem paired nicking seems to accomplish targeted knock-in via DNA recombination analogous to Holliday’s model, and creates intended genetic changes in the genome without introducing additional nucleotide changes such as silent mutations. Targeted knock-in through tandem paired nicking neither triggers significant p53 activation nor occurs preferentially in p53-suppressed cells. These properties of tandem paired nicking demonstrate its utility in precision genome engineering.