TY - JOUR
T1 - Robust and efficient knock-in in embryonic stem cells and early-stage embryos of the common marmoset using the CRISPR-Cas9 system
AU - Yoshimatsu, Sho
AU - Okahara, Junko
AU - Sone, Takefumi
AU - Takeda, Yuta
AU - Nakamura, Mari
AU - Sasaki, Erika
AU - Kishi, Noriyuki
AU - Shiozawa, Seiji
AU - Okano, Hideyuki
N1 - Funding Information:
Some data presented herein were obtained during the “Construction of System for Spread of Primate Model Animals” project, supported by the Strategic Research Program for Brain Sciences and Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) of the MEXT and the AMED (to H.O. and S. S., E. S); and Scientific Research in Innovative Areas, a MEXT Grant-in-Aid project FY2014-2018 “Brain Protein Aging and Dementia Control” (to H.O.). This work was also supported by RIKEN Junior Research Associate Program (to S.Y.). We thank Eiki Takahashi (RIKEN) and Sumitomo Dainippon Pharma Co., Ltd. for kindly providing experimental materials.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85061156432&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85061156432&partnerID=8YFLogxK
U2 - 10.1038/s41598-018-37990-w
DO - 10.1038/s41598-018-37990-w
M3 - Article
C2 - 30728412
AN - SCOPUS:85061156432
SN - 2045-2322
VL - 9
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 1528
ER -