Abstract
Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) (CRISPR-Cas9) gene editing enables rapid production of genome-edited animals. The Cas9/guide RNA (gRNA) component can be introduced into zygotes in several ways. Here, we provide an instructional guide for the generation of knockout mice using cytoplasmic injection of in vitro transcribed Cas9 RNA and gRNA.
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References
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Acknowledgments
We thank Dr. Yuji Arai (National Cerebral and Cardiovascular Center) for technical advice on microinjection and Mika Kimura (Gunma University) for proofreading the section describing in vitro transcription of RNA. This work was supported by grants from the Basic Science and Platform Technology Program for Innovative Biological Medicine from the Ministry of Education, Culture, Sports, Science, and Technology, Japan (MEXT); the Research Support Project for Life Science and Drug Discovery (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) from Agency for Medical Research and Development (AMED) under Grant Number JP22ama121049; and the Practical Research Project for Rare/Intractable Diseases from AMED.
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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Horii, T., Hatada, I. (2023). Generation of Genome-Edited Mice by Cytoplasmic Injection of CRISPR-Cas9 RNA. In: Hatada, I. (eds) Genome Editing in Animals. Methods in Molecular Biology, vol 2637. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3016-7_6
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DOI: https://doi.org/10.1007/978-1-0716-3016-7_6
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3015-0
Online ISBN: 978-1-0716-3016-7
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