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

Sho Yoshimatsu, Junko Okahara, Takefumi Sone, Yuta Takeda, Mari Nakamura, Erika Sasaki, Noriyuki Kishi, Seiji Shiozawa, Hideyuki Okano

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish
Article number1528
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 2019 Dec 1

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Clustered Regularly Interspaced Short Palindromic Repeats
Callithrix
Embryonic Stem Cells
Embryonic Structures
Homologous Recombination
Double-Stranded DNA Breaks
Primates
Biomedical Research

ASJC Scopus subject areas

  • General

Cite this

Robust and efficient knock-in in embryonic stem cells and early-stage embryos of the common marmoset using the CRISPR-Cas9 system. / Yoshimatsu, Sho; Okahara, Junko; Sone, Takefumi; Takeda, Yuta; Nakamura, Mari; Sasaki, Erika; Kishi, Noriyuki; Shiozawa, Seiji; Okano, Hideyuki.

In: Scientific Reports, Vol. 9, No. 1, 1528, 01.12.2019.

Research output: Contribution to journalArticle

Yoshimatsu, Sho ; Okahara, Junko ; Sone, Takefumi ; Takeda, Yuta ; Nakamura, Mari ; Sasaki, Erika ; Kishi, Noriyuki ; Shiozawa, Seiji ; Okano, Hideyuki. / Robust and efficient knock-in in embryonic stem cells and early-stage embryos of the common marmoset using the CRISPR-Cas9 system. In: Scientific Reports. 2019 ; Vol. 9, No. 1.
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AU - Nakamura, Mari

AU - Sasaki, Erika

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