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

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


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Callithrix
  • DNA Breaks, Double-Stranded*
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / metabolism
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Female
  • Forkhead Transcription Factors / antagonists & inhibitors
  • Forkhead Transcription Factors / genetics
  • Gene Editing*
  • Gene Knock-In Techniques / methods*
  • Gene Targeting
  • Homologous Recombination
  • Humans
  • Male
  • Models, Animal
  • Myelin Proteolipid Protein / antagonists & inhibitors
  • Myelin Proteolipid Protein / genetics
  • Neural Stem Cells / cytology*
  • Neural Stem Cells / metabolism


  • FOXP2 protein, human
  • Forkhead Transcription Factors
  • Myelin Proteolipid Protein