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A Transgenic Core Facility’s Experience in Genome Editing Revolution

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Book cover Precision Medicine, CRISPR, and Genome Engineering

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1016))

Abstract

The use of animal models, particularly rodents, has been immensely important to nearly all aspects of biomedical research, from basic science exploration to translational discoveries into clinical applications. The transgenic core facility that provides animal model production, preservation, and recovery services has been fundamental to the success of research efforts using animals. Recent advances in genome editing technologies, especially the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) enzyme system, have transformed the tedious animal model production into a simple and effective procedure. We, as a transgenic core facility established in 1993, adopted the CRISPR/Cas9 technology in early 2014 and have experienced the dramatic shift in the practice of animal model production, from the conventional embryonic stem cell approach to the direct genomic editing in rodent embryos. In this chapter, we describe the lessons that we learned from more than 200 genome editing projects performed in this core facility within the past 3 years. We also provide the practical guidelines for efficient generation of animal models using this technology and the insights into where new technologies lead us.

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Abbreviations

Cas9:

CRISPR associated protein 9

CRISPR:

Clustered regularly interspaced short palindromic repeats

ES cells:

Embryonic stem cells

HDR:

Homology-directed repair

NHEJ:

Non-homologous end joining

sgRNA:

Single-guide RNA

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Acknowledgements

We thank current and former staff members, Yinhuai Chen, Huirong Xie, Alexandra Falcone, Susan Martin, Melissa Scott, Evan Barr-Beare, Calista Falcone, Kendall Smith and Kristen Martin for performing the services and generating the data for this article; Melissa Scott for commenting on the manuscript. This work was supported by Cincinnati Children’s Research Foundation.

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Authors and Affiliations

  1. Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center, 3333 Burnet Avenue, MLC7007, Cincinnati, OH, 45229, USA

    Celvie L. Yuan Ph.D. & Yueh-Chiang Hu Ph.D.

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  1. Celvie L. Yuan Ph.D.
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  2. Yueh-Chiang Hu Ph.D.
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Correspondence to Yueh-Chiang Hu Ph.D. .

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Editors and Affiliations

  1. Departments of Ophthalmology, Pathology & Cell Biology and Biomedical Engineering, Institute of Human Nutrition, Columbia Stem Cell Initiative, Herbert Irving Comprehensive Cancer Center, Graduate Program in Neurobiology & Behavior, Columbia University, New York, New York, USA

    Stephen H. Tsang

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Yuan, C.L., Hu, YC. (2017). A Transgenic Core Facility’s Experience in Genome Editing Revolution. In: Tsang, S. (eds) Precision Medicine, CRISPR, and Genome Engineering. Advances in Experimental Medicine and Biology, vol 1016. Springer, Cham. https://doi.org/10.1007/978-3-319-63904-8_4

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