Abstract
Humane use of preclinical large animal cancer models plays a critical role in understanding cancer biology and developing therapeutic treatments. Among the large animal candidates, goats have great potentials as sustainable sources for large animal cancer model development. Goats are easier to handle and cheaper to raise. The genome of the goats has been sequenced recently. It has been known that goats develop skin, adrenal cortex, breast and other types of cancers. Technically, goats are subject to somatic cell nuclear transfer more efficiently and exhibit better viability through the cloning process. Towards the development of a goat cancer model, we created a transgenic goat fetal fibroblast (GFF) cell as the donor cell for SCNT. Human mutated K-ras (hK-rasG12D) was chosen as the transgene, as it is present in 20% of cancers. Both hK-rasG12D and a herpes simplex viral thymidine kinase (HSV1-tk) reporter genes, flanked by a pair of LoxP sites, were knocked in the GFF endogenous K-ras locus through homologous recombination. Following Cre-mediated activation (with a 95% activation efficiency), hK-rasG12D and HSV1-tk were expressed in the transgenic GFF cells, evidently through the presence of corresponding mRNAs, and confirmed by HSV1-tk protein function assay. The hK-rasG12D expressing GFF cells exhibited enhanced proliferation rates and an anchorage-independent growth behavior. They were able to initiate tumor growth in athymic nude mice. In conclusion, after activating hK-rasG12D gene expression, hK-rasG12D transgenic GFF cells were transformed into tumorgenesis cells. Transgenic goats via SCNT using the above-motioned cells as the donor cells have been established.
Highlights
Mice are the most commonly used animal model due to the vast array of reagents and gene manipulation strategies currently available for this species
We developed transgenic goat cells with hK-ras and HSV1-tk genes using primary goat fetal fibroblast (GFF) cells
Comparing with similar works on mouse model development [12,13,14,15], we found wild-type GFF cells are quite different from mouse embryonic fibroblast (MEF) cells
Summary
Mice are the most commonly used animal model due to the vast array of reagents and gene manipulation strategies currently available for this species. The small size of the mouse and its anatomical structures present problematic issues when measuring the pathophysiologic parameters of cancer or other diseases. This is especially evident when comparing the vastly different physiologic values between mice and humans. The choice of goats for cancer model development is motivated by 1) goats develop skin, adrenal cortex, breast and other types of cancers [1,2,3,4]; 2) goat genome has been sequenced recently [5]; and 3) comparing to pigs or other large animals, goats are more robust to the cloning process, easier to handle and less expensive to raise, which makes them sustainable as an animal model source
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.