Journal
BIOTECHNOLOGY JOURNAL
Volume 17, Issue 7, Pages -Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/biot.202100434
Keywords
CRISPR-Cas9; pig; somatic cell nuclear transfer; transgenic cancer model
Funding
- National Research Foundation of Korea [NRF-2017R1A2B4002546]
- Global Research and Development Center (GRDC) Program
- Ministry of Education, Science and Technology [2017K1A4A3014959]
- Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET)
- Agri-Bio industry Technology Development Program
- Ministry of Agriculture, Food and Rural Affairs (MAFRA) [318016-5]
- Institute of Animal Molecular Biotechnology Grant
- School of Life Sciences and Biotechnology for BK21 Plus, Korea University
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In this study, a new onco-pig model was successfully developed using CRISPR-Cas9 technology, which exhibited cancer characteristics and could be used for cancer research. The results of the study indicate that this model has a wide range of potential applications.
Alternative cancer models that are close to humans are required to create more valuable preclinical results during oncology studies. Here, a new onco-pig model via developing a CRISPR-Cas9-based Conditional Polycistronic gene expression Cassette (CRI-CPC) system to control the tumor inducing simian virus 40 large T antigen (SV40LT) and oncogenic HRAS(G12V). After conducting somatic cell nuclear transfer (SCNT), transgenic embryos were transplanted into surrogate mothers and five male piglets were born. Umbilical cord analysis confirmed that all piglets were transgenic. Two of them survived and they expressed a detectable green fluorescence. The test was made whether CRI-CPC models were naturally fertile and whether the CRI-CPC system was stably transferred to the offspring. By mating with a normal female pig, four offspring piglets were successfully produced. Among them, only three male piglets were transgenic. Finally, their applicability was tested as cancer models after transduction of Cas9 into fibroblasts from each CRI-CPC pig in vitro, resulting in cell acquisition of cancerous characteristics via the induction of oncogene expression. These results showed that our new CRISPR-Cas9-based onco-pig model was successfully developed.
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