学術雑誌論文

学術雑誌論文

Scientific Reports. 2016, 6, p. 31666

CRISPR/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice

Nature Research

英語

2016-08-17

https://doi.org/10.1038/srep31666

Oji, A., Noda, T., Fujihara, Y. et al. CRISPR/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice. Sci Rep 6, 31666 (2016). https://doi.org/10.1038/srep31666

Targeted gene disrupted mice can be efficiently generated by expressing a single guide RNA (sgRNA)/CAS9 complex in the zygote. However, the limited success of complicated genome editing, such as large deletions, point mutations, and knockins, remains to be improved. Further, the mosaicism in founder generations complicates the genotypic and phenotypic analyses in these animals. Here we show that large deletions with two sgRNAs as well as dsDNA-mediated point mutations are efficient in mouse embryonic stem cells (ESCs). The dsDNA-mediated gene knockins are also feasible in ESCs. Finally, we generated chimeric mice with biallelic mutant ESCs for a lethal gene, Dnajb13, and analyzed their phenotypes. Not only was the lethal phenotype of hydrocephalus suppressed, but we also found that Dnajb13 is required for sperm cilia formation. The combination of biallelic genome editing in ESCs and subsequent chimeric analysis provides a useful tool for rapid gene function analysis in the whole organism.

© 2016 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License.

78585

CRISPR/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice

Nature Research

英語

2016-08-17

Scientific Reports

6

31666

https://doi.org/10.1038/srep31666

27530713

https://hdl.handle.net/11094/78585