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Simple, Efficient CRISPR-Cas9-Mediated Gene Editing in Mice: Strategies and Methods

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Mouse Models for Drug Discovery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1438))

Abstract

Genetic modification of almost any species is now possible using approaches based on targeted nucleases. These novel tools now bypass previous limited species windows, allowing precision nucleotide modification of the genome at high efficiency, rapidly and economically. Here we focus on the modification of the mouse genome; the mouse, with its short generation time and comparatively low maintenance/production costs is the perfect mammal with which to probe the genome to understand its functions and complexities. Further, using targeted nucleases combined with homologous recombination, it is now possible to precisely tailor the genome, creating models of human diseases and conditions directly and efficiently in zygotes derived from any mouse strain. Combined these approaches make it possible to sequentially and progressively refine mouse models to better reflect human disease, test and develop therapeutics. Here, we briefly review the strategies involved in designing targeted nucleases (sgRNAs) providing solutions and outlining in detail the practical processes involved in precision targeting and modification of the mouse genome and the establishing of new precision genetically modified mouse lines.

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Acknowledgements

This work was done in close cooperation with The Jackson Laboratory microinjection team. We also fully acknowledge the help and excellent assistance of Cindy Avery and Deb Woodworth for genotyping and animal maintenance, plus discussions regarding CRISPR with Drs. Vishnu Hosur, Wenning Qin, and Gabriele Proetzel. Funding for this work was provided by the National Institutes of Health Grant OD011190 (M.V.W.) and The Jackson Laboratory. We also give our thanks to Addgene for supplying many of the plasmids used in our studies.

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  1. The Jackson Laboratory, 600 Main Street, Bar Harbor, ME, 04609-1500, USA

    Benjamin E. Low, Peter M. Kutny & Michael V. Wiles

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  1. Benjamin E. Low
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  2. Peter M. Kutny
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Correspondence to Michael V. Wiles .

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  1. Takeda Pharmaceutical Company, Cambridge, Massachusetts, USA

    Gabriele Proetzel

  2. The Jackson Laboratory, Bar Harbor, Maine, USA

    Michael V. Wiles

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Low, B.E., Kutny, P.M., Wiles, M.V. (2016). Simple, Efficient CRISPR-Cas9-Mediated Gene Editing in Mice: Strategies and Methods. In: Proetzel, G., Wiles, M. (eds) Mouse Models for Drug Discovery. Methods in Molecular Biology, vol 1438. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3661-8_2

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