© Springer International Publishing AG 2017 75
S.H. Tsang (ed.), Precision Medicine, CRISPR, and Genome Engineering,
Advances in Experimental Medicine and Biology 1016,
DOI 10.1007/978-3-319-63904-8_4
Chapter 4
A Transgenic Core Facility’s Experience
in Genome Editing Revolution
Celvie L. Yuan and Yueh-Chiang Hu
Abstract The use of animal models, particularly rodents, has been immensely
important to nearly all aspects of biomedical research, from basic science explora-
tion to translational discoveries into clinical applications. The transgenic core facil-
ity 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 proce-
dure. 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 prac-
tice 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.
Keywords CRISPR/Cas9 • Microinjection • Transgenic facility • Animal model •
Genome editing • sgRNA
Abbreviations
Cas9 CRISPR associated protein 9
CRISPR Clustered regularly interspaced short palindromic repeats
ES cells Embryonic stem cells
HDR Homology-directed repair
C.L. Yuan, Ph.D. • Y.-C. Hu, Ph.D. (*)
Division of Developmental Biology, Cincinnati Children’s Hospital Medical Center,
3333 Burnet Avenue, MLC7007, Cincinnati, OH 45229, USA
e-mail: [email protected]; [email protected]