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

Non-human primates (NHPs) are the closest animal model to humans; thus, gene engineering technology in these species holds great promise for the elucidation of higher brain functions and human disease models. Knockin (KI) gene targeting is a versatile approach to modify gene(s) of interest; however, it generally suffers from the low efficiency of homology-directed repair (HDR) in mammalian cells, especially in non-expressed gene loci. In the current study, we generated a tyrosine hydroxylase (TH)-2A-Cre KI model of the common marmoset monkey (marmoset; Callithrix jacchus) using an HDR-biased CRISPR-Cas9 genome editing approach using Cas9-DN1S and RAD51. This model should enable labeling and modification of a specific neuronal lineage using the Cre-loxP system. Collectively, the current study paves the way for versatile gene engineering in NHPs, which may be a significant step toward further biomedical and preclinical applications.
Competing Interests: Declaration of interests H.O. has been a paid scientific advisory board member of San Bio Co. Ltd., Regenerative Medicine iPS Gateway Center Co. Ltd., and K Pharma, Inc. S.Y. has been a paid associate researcher of Daiichi-Sankyo RD Novare Co. Ltd. However, there was no effect of these companies on the interpretation, writing, or publication of this study. H.O. and S.Y. declare that there are no non-financial conflicts of interest with this work. In addition, the other authors declare that there are neither financial nor non-financial conflicts of interest.
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