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Embryo-Based Large Fragment Knock-in in Mammals: Why, How and What’s Next

by Steven Erwood 1,2 and Bin Gu 3,*
1
Program in in Genetics and Genome Biology, Hospital for Sick Children, Toronto, ON M5G 0A4, Canada
2
Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada
3
Program in Developmental and Stem Cell Biology, Hospital for Sick Children, Toronto, ON M5G 0A4, Canada
*
Author to whom correspondence should be addressed.
Genes 2020, 11(2), 140; https://doi.org/10.3390/genes11020140
Received: 31 December 2019 / Revised: 24 January 2020 / Accepted: 26 January 2020 / Published: 29 January 2020
(This article belongs to the Special Issue Prospects in Transgenic Technology 2020)
Endonuclease-mediated genome editing technologies, most notably CRISPR/Cas9, have revolutionized animal genetics by allowing for precise genome editing directly through embryo manipulations. As endonuclease-mediated model generation became commonplace, large fragment knock-in remained one of the most challenging types of genetic modification. Due to their unique value in biological and biomedical research, however, a diverse range of technological innovations have been developed to achieve efficient large fragment knock-in in mammalian animal model generation, with a particular focus on mice. Here, we first discuss some examples that illustrate the importance of large fragment knock-in animal models and then detail a subset of the recent technological advancements that have allowed for efficient large fragment knock-in. Finally, we envision the future development of even larger fragment knock-ins performed in even larger animal models, the next step in expanding the potential of large fragment knock-in in animal models.
Keywords: CRISPR-Cas9; genome editing; large fragment knock-in; HDR; embryo CRISPR-Cas9; genome editing; large fragment knock-in; HDR; embryo
MDPI and ACS Style

Erwood, S.; Gu, B. Embryo-Based Large Fragment Knock-in in Mammals: Why, How and What’s Next. Genes 2020, 11, 140.

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