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Delivery of CRISPR-Cas9 into Mouse Zygotes by Electroporation

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Microinjection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1874))

Abstract

The CRISPR-Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including the mice. In this scheme, components of the CRISPR-Cas9 system are delivered into the mouse zygote and mutant mice carrying genetic modifications derived. Although microinjection has been the technology of choice, electroporation has also emerged and been proven to be effective delivering CRISPR-Cas9 reagents into the mouse zygote. Here, we describe the experimental protocol employing electroporation to deliver CRISPR-Cas9 reagents into mouse embryos and derive gene-edited mutant mice.

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Acknowledgment

We would like to thank the Genetic Engineering Technologies and the Reproductive sciences groups of the Jackson Laboratory for their partnership and contribution exploring into the CRISPR-Cas9 technology. Research reported in this publication was partially supported by the National Cancer Institute under award number P30CA034196. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. H.W. is supported by the Strategic Priority Research Program of the Chinese Academy of Sciences (No. XDA16010205), “National Natural Science Foundation of China” (31471215), and the National Key Research and Development Program of China (No. 2016YFA0101402).

Author information

Authors and Affiliations

  1. The Jackson Laboratory, Bar Harbor, ME, USA

    Wenning Qin

  2. State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China

    Haoyi Wang

  3. Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China

    Haoyi Wang

Authors
  1. Wenning Qin
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  2. Haoyi Wang
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Editor information

Editors and Affiliations

  1. National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Chengyu Liu

  2. National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Yubin Du

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Qin, W., Wang, H. (2019). Delivery of CRISPR-Cas9 into Mouse Zygotes by Electroporation. In: Liu, C., Du, Y. (eds) Microinjection. Methods in Molecular Biology, vol 1874. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8831-0_10

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  • DOI: https://doi.org/10.1007/978-1-4939-8831-0_10

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8830-3

  • Online ISBN: 978-1-4939-8831-0

  • eBook Packages: Springer Protocols

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