Generation of a tyrosine hydroxylase-2A-Cre knockin non-human primate model by homology-directed-repair-biased CRISPR genome editing

• DOI: 10.1016/j.crmeth.2023.100590
• Publication Date: 2023-09-15T10:52:29Z
• Authors: Sho Yoshimatsu; Junko Okahara; Junko Yoshie; Yoko Igarashi; Ryusuke Nakajima; Tsukasa Sanosaka; Emi Qian; Tsukika Sato; Hiroya Kobayashi; Satoru Morimoto; Noriyuki Kishi; Devin M. Pillis; Punam Malik; Toshiaki Noce; Hideyuki Okano
• Keywords: Homology

Non-human primates (NHPs) are the closest animal model to humans; thus, gene engineering technology in these species holds great promise for elucidation of higher brain functions and human disease models. Knockin (KI) targeting is a versatile approach modify gene(s) interest; however, it generally suffers from low efficiency homology-directed repair (HDR) mammalian cells, especially non-expressed loci. In current study, we generated tyrosine hydroxylase (TH)-2A-Cre KI common marmoset monkey (marmoset; Callithrix jacchus) using an HDR-biased CRISPR-Cas9 genome editing Cas9-DN1S RAD51. This should enable labeling modification specific neuronal lineage Cre-loxP system. Collectively, study paves way NHPs, which may be significant step toward further biomedical preclinical applications.