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Sci Rep. 2019 Feb 6;9(1):1528. doi: 10.1038/s41598-018-37990-w.

Robust and efficient knock-in in embryonic stem cells and early-stage embryos of the common marmoset using the CRISPR-Cas9 system.

Author information

1
Department of Physiology, School of Medicine, Keio University, Shinjuku-ku, Tokyo, 160-8582, Japan.
2
Laboratory for Proteolytic Neuroscience, RIKEN Center for Brain Science, Wako City, Saitama, 351-0198, Japan.
3
Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako City, Saitama, 351-0198, Japan.
4
Central Institute for Experimental Animals, Kawasaki, Kanagawa, 210-0821, Japan.
5
Department of Physiology, School of Medicine, Keio University, Shinjuku-ku, Tokyo, 160-8582, Japan. shiozawa@a7.keio.jp.
6
Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako City, Saitama, 351-0198, Japan. shiozawa@a7.keio.jp.
7
Department of Physiology, School of Medicine, Keio University, Shinjuku-ku, Tokyo, 160-8582, Japan. hidokano@a2.keio.jp.
8
Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, Wako City, Saitama, 351-0198, Japan. hidokano@a2.keio.jp.

Abstract

Genome editing technology greatly facilitates the genetic modification of various cells and animals. The common marmoset (Callithrix jacchus), a small non-human primate which exhibits high reproductive efficiency, is a widely used animal model in biomedical research. Developing genome editing techniques in the common marmoset will further enhance its utility. Here, we report the successful establishment of a knock-in (KI) method for marmoset embryonic stem cells (ESCs), which is based on the CRISPR-Cas9 system. The use of CRISPR-Cas9, mediated by homologous recombination (HR), enhanced the KI efficiency in marmoset ESCs. Furthermore, we succeeded in performing KI in early-stage marmoset embryos. In the course of the experiments, we found that HR in the marmoset ESCs is innately highly efficient. This suggested that the marmoset possesses a repair mechanism for DNA double-strand breaks. The current study will facilitate the generation of genetically modified marmosets and gene function analysis in the marmoset.

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