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Methodology for Efficient and Biosecure Genetic Engineering of Biomedical Swine Models
Author(s) -
Meudt Jennifer J.,
Reichert Jamie L.,
Lopez Ana Cecilia Escobar,
Krentz Kathleen J.,
Rubinstein Clifford Dustin,
Lehman Brent P.,
Schomberg Dominic T.,
Monson Ricky L.,
Willenburg Kilby,
Frank Jennifer M.,
Graff Keri D.,
Shanmuganayagam Dhanansayan
Publication year - 2020
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2020.34.s1.06874
Subject(s) - biosecurity , workflow , crispr , genetic model , computer science , data science , computational biology , biology , genetics , gene , ecology , database
The genetic proximity of swine to human, the overwhelming anatomical, physiological and pathophysiological similarities, and the ability to mimic external factors in a practical manner make swine the ideal preclinical model for many human diseases. While the value of swine in biomedical research is well recognized, their use has been limited by the lack of tools to easily edit the swine genome to model human disease, and the lack of facilities, resources and expertise at most institutions for supporting large‐scale swine research. The recent implementation of new genetic editing tools such as the CRISPR/Cas9 platform make the creation of novel genetic swine models possible and practical. However, conventional approaches to genetic engineering of swine rely on ova obtained from commercial abattoirs, significantly compromising biosecurity, and often offering little control over the background genetic of the swine breeds upon which the genomic editing is performed. Here, we describe methods and workflow that we have developed that allow for efficient genetic engineering of swine, while maintaining control over background genetics and minimizing the biosecurity risk to the animals and the facility. The process centrally relies on the refinement of reproductive cycle synchronization and superovulation of swine and time‐precise coordination of the entire genetic engineering workflow. Support or Funding Information The research was supported by the Biomedical & Genomic Research Group Discretionary Fund (University of Wisconsin‐Madison), NF North Central, NF Network, and NF Team Foundation