Abstract
In mammalian cells, genome editing with the single guide RNA (sgRNA)/Cas9 complex allows for high targeting efficiency within a relatively short time frame with the added benefits of being low cost and easy to design. sgRNA/Cas9-mediated editing in mouse zygotes has accelerated the analysis of gene functions and the generation of mouse models of human diseases. Despite the benefits, this method still suffers from several problems, such as mosaicism in the founder generation which complicates genotyping and phenotypical analyses, and the low efficiency of more complicated genome editing. Thus, we recently established the system for genome editing in embryonic stem (ES) cells and its application for chimeric analysis in mice. In this section, we introduce the procedure for genome editing in mouse zygotes and ES cells.
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Acknowledgment
This work was supported by the MEXT/JSPS KAKENHI grants (JP16J04613 to T.N., JP15J04519 to A.O., and JP25112007 and JP25250014 to M.I.), and the Takeda Science Foundation grant to M.I. The authors would like to thank Dr. Julio Castaneda for critical reading of the manuscript.
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Noda, T., Oji, A., Ikawa, M. (2017). Genome Editing in Mouse Zygotes and Embryonic Stem Cells by Introducing SgRNA/Cas9 Expressing Plasmids. In: Hatada, I. (eds) Genome Editing in Animals. Methods in Molecular Biology, vol 1630. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7128-2_6
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DOI: https://doi.org/10.1007/978-1-4939-7128-2_6
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