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Production of transgenic pigs using the pGFAP-CreERT2;EGFPLoxP inducible system for central nervous system disease models
Seon-Ung Hwang1, 2,†, Kiyoung Eun3,†, Junchul David Yoon1,2, Hyunggee Kim3,*, Sang-Hwan Hyun1,2,*
1Laboratory of Veterinary Embryology and Biotechnology, Veterinary Medical Center and College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea
2Institute of Stem Cell & Regenerative Medicine, Chungbuk National University, Cheongju, Korea
3Department of Biotechnology, School of Life Sciences and Biotechnology, Korea University, Seoul, Korea
Correspondence to: Tel: +82-43-261-3393; Fax: +82-43-267-3150; E-mail: hg-kim@korea.ac.kr (H Kim), shhyun@cbu.ac.kr (SH Hyun)
The first two authors contributed equally to this work.
Received: July 24, 2017; Revised: October 1, 2017; Accepted: November 24, 2017; Published online: December 28, 2017.
Transgenic (TG) pigs are an important model for biomedical research, including disease modeling, pharmaceutical toxicity testing, and regenerative medicine. Here, we constructed 2 vector systems using the promoter of the pig glial fibrillary acidic protein (pGFAP) gene, which is an astrocyte cell marker. We established donor TG fibroblasts with pGFAP-CreERT2; LCMV-EGFPLoxP. We evaluated the effect of the transgenes on TG-SCNT embryo development. The cleavage rates were not significantly different among control and transgene-donor group. Next, embryo transfer was performed thrice just before the ovulation of surrogate sows. One sow delivered 5 TG piglets at 115 days after pregnancy. Polymerase chain reaction (PCR) analysis with genomic DNA isolated from skin tissues of TG pigs revealed that all 5 TG pigs had the transgenes. EGFP expression in all organs tested was confirmed by immunofluorescence staining and PCR. Real-time PCR analysis showed that pGFAP promoter-driven Cre fused to the mutated human ligand-binding domain of the estrogen receptor (CreERT2) mRNA was highly expressed in the cerebrum. Semi-nested PCR analysis revealed that CreERT2-mediated recombination was induced in the cerebrum and cerebellum but not in the skin. Thus, we successfully generated a TG pig with a 4-hydroxytamoxifen (TM)-inducible pGFAP-CreERT2;EGFPLoxP recombination system by somatic cell nuclear transfer (SCNT).
Keywords: Porcine, GFAP, CreERT2, SCNT, Transgenic pigs

© 2017 The Korean Society of Veterinary Science.