Abstract
CRISPR/Cas enables rapid production of genome-edited animals. The Cas9/gRNA component can be introduced to fertilized eggs in several ways. Here, we provide an instructional guide for the generation of knockout mice by cytoplasmic injection using in vitro transcribed Cas9 and gRNA.
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Acknowledgment
We thank Dr. Yuji Arai (National Cerebral and Cardiovascular Center) for technical advice on microinjection, and Mika Kimura (Gunma University) for proofreading of the section describing in vitro transcription of RNA. This work was supported by the Basic Science and Platform Technology Program for Innovative Biological Medicine from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (MEXT) and the Agency for Medical Research and Development (AMED) of Japan.
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Horii, T., Hatada, I. (2017). Genome Editing of Mouse by Cytoplasmic Injection. 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_5
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DOI: https://doi.org/10.1007/978-1-4939-7128-2_5
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7127-5
Online ISBN: 978-1-4939-7128-2
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