ORIGINAL RESEARCH article
Front. Genome Ed.
Sec. Genome Editing Tools and Mechanisms
This article is part of the Research TopicRecent Advances in Genome Editing Technology and Its Multifaceted Applications in Medicine, Agriculture, and CosmeticsView all articles
The precision strategy of human genome correction via a set of circular donor-DNA and its cleaver
Provisionally accepted
- ID Pharma Co., Ltd., Tsukuba, Japan
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Homologous recombination (HR) corrects a mutational sequence causing a genetic disease with the normal sequence so as to recover healthy state in humans. A targeted genomic breakage such as CRISPR-Cas9 can induce a copy-paste type HR event; however, CRISPR-Cas9 more frequently induces imprecise non-homologous end-joining events, leading to one-step multiple knockout products for paralogous genes or homologous alleles as a special merit. We have established the precision strategy of the crossover type HR-based gene editing that is primed by intra-cellular circular donor cleavage (InCDC). The InCDC technique generates targeted duplication of the circular donor-plasmid at the target locus in human cells to form a doublet configuration comprising the donor DNA with the designed sequence and the target DNA with the original sequence, with much higher efficiency than conventional donor-linearization techniques. This doublet form leads to the singlet form remaining the designed allele. We found that the safety distance within designed circular donor-plasmid and its intra-cellular cleavage were particularly critical to protect a designed sequence from enzymatic exclusion and propose that the InCDC technology leads to precision genome editing, such as the replacement of the genetic disease-causative allele with the designed correct allele.
Keywords: Crossover type Homologous recombination, Intra-cellular circular donor cleavage, Natural replacement, Safety distance, Targeted duplication
Received: 03 Oct 2025; Accepted: 22 Jan 2026.
Copyright: © 2026 Kusano, Takizawa, Kawaguchi, Hara and Mori. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Kohji Kusano
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