High-efficiency homology-directed insertion into the genome using the engineered homing endonuclease ARCUS.

ABSTRACT: Several gene editing tools have entered the clinic, representing varied options for eliminating or correcting mutations. Although gene editing by homologous recombination (HR) can potentially accomplish any type of gene edit (insertions, deletions, and replacements), as the outcome is defined by a recombinant repair template, gene editing enzymes that support efficient HR are rare. ARCUS nucleases, engineered from the homing endonuclease I-CreI, have programmable sequence specificity and support precise, high-frequency transgene insertion. In this study, we demonstrate that the 3' overhangs that ARCUS nucleases generate when cutting DNA are key to triggering high rates of HR. We show that a single editor can be used to accomplish the full range of currently understood DNA editing approache

SUBMITTER: Resly LC 

PROVIDER: S-EPMC12497482 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature