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Using Microinjection to Generate Genetically Modified Caenorhabditis elegans by CRISPR/Cas9 Editing

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Microinjection

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

Abstract

In this chapter, we describe the procedure for generating genetically modified Caenorhabditis elegans using microinjection via the Cas9-mediated Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) editing technique. Specifically, we describe the detailed method of performing CRISPR editing by microinjection using the Cloning-free Co-CRISPR method described by the Seydoux lab. This microinjection protocol can also be used for CRISPR editing with protocols from other labs as well as for a variety of other editing techniques including Mos1-mediated single-copy transgene insertions (MosSCI), transcriptional activator-like nucleases (TALENs), and zinc-finger nucleases (ZFNs). Further, this microinjection protocol can also be used for injecting plasmid DNA to generate heritable extrachromosomal arrays for gene expression and mosaic analysis, performing RNAi by injection and delivering RNA, dyes or other molecules into the C. elegans germline.

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Acknowledgments

We would like to thank Kevin O’Connell, Andy Golden, Aimee Jaramillo-Lambert, Tetsu Fukushige, Prabhu Sankaralingam, and Daman Kumari from the National Institutes of Health for their critical feedback and comments on this manuscript. Nicole DeVaul and Tyler Hansen contributed equally to this chapter.

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Author notes

  1. Nicole DeVaul and Tyler Hansen contributed equally to this chapter.

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, The University of Tulsa, Tulsa, OK, USA

    Jyoti Iyer

  2. The Department of Anatomy and Cell Biology, The George Washington University School of Medicine and Health Sciences, Tulsa, OK, USA

    Nicole DeVaul

  3. Department of Biochemistry, Vanderbilt University School of Medicine, Tulsa, OK, USA

    Tyler Hansen

  4. Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, Tulsa, OK, USA

    Ben Nebenfuehr

Authors
  1. Jyoti Iyer
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  2. Nicole DeVaul
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  3. Tyler Hansen
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  4. Ben Nebenfuehr
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Corresponding author

Correspondence to Jyoti Iyer .

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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© 2019 Springer Science+Business Media, LLC, part of Springer Nature

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Iyer, J., DeVaul, N., Hansen, T., Nebenfuehr, B. (2019). Using Microinjection to Generate Genetically Modified Caenorhabditis elegans by CRISPR/Cas9 Editing. 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_25

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  • DOI: https://doi.org/10.1007/978-1-4939-8831-0_25

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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

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