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Construction of transgenic swine with induced expression of Cre recombinase

Published online by Cambridge University Press:  07 January 2010

L. Chen
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China The Basis of the Faculty of Medicine, Department of BeiHua University, Jilin 132013, PR China
L. Li
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
D. Pang
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
Z. Li
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
T. Wang
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
M. Zhang
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
N. Song
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
S. Yan
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
L. X. Lai
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
H. Ouyang*
Affiliation:
Department of Animal Biotechnology, College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun 130062, PR China
*
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Abstract

Miniature pigs have been recognized as valuable experimental animals in medical research. However, porcine models related to gene knockout of human diseases are not widely available. The objective of this study was to establish Mx1-Cre pigs using somatic cell nuclear transfer. In this study, we created transgenic pigs using somatic cell nuclear transfer (SCNT). Transfer of 210, 230, 250 and 215 zygotes to four surrogates produced 10 piglets. The Cre recombinase expression in transgenic pigs was studied using reverse transcriptase (RT)-PCR and immunohistochemistry. Mx1-Cre swine were shown to harbor the Cre gene in their genomic DNA using the PCR. In conclusion, Mx1-Cre transgenic piglets were successfully produced by SCNT. These transgenic swine, in conjunction with inducible systems for controlling Cre expression and function, are likely to have a profound impact on the study of human diseases.

Type
Full Paper
Copyright
Copyright © The Animal Consortium 2009

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Footnotes

a

L. Chen and L. Li contributed equally to this work

References

Chen, M, Wolfe, A, Wang, X, Chang, C, Yeh, S, Radovick, S 2009. Generation and characterization of a complete null estrogen receptor alpha mouse using Cre/LoxP technology. Molecular and Cellular Biochemistry 321, 145153.CrossRefGoogle ScholarPubMed
Doetschman, T, Gregg, RG, Maeda, N, Hooper, ML, Melton, DW, Thompson, S, Smithies, O 1987. Targetted correction of a mutant HPRT gene in mouse embryonic stem cells. Nature 330, 576578.CrossRefGoogle ScholarPubMed
Forsberg, EJ, Strelchenko, NS, Augenstein, ML, Betthauser, JM, Childs, LA, Eilertsen, KJ, Enos, JM, Forsythe, TM, Golueke, PJ, Koppang, RW, Lange, G, Lesmeister, TL, Mallon, KS, Mell, GD, Misica, PM, Pace, MM, Pfister-Genskow, M, Voelker, GR, Watt, SR, Bishop, MD 2002. Production of cloned cattle from in vitro systems. Biology of Reproduction 67, 327333.CrossRefGoogle ScholarPubMed
Fujimura, T, Murakami, H, Kurome, M, Takahagi, Y, Shigehisa, T, Nagashima, H 2008. Effects of recloning on the efficiency of production of 1,3-galactosyltransferase-knockout pigs. The Journal of Reproduction and Development 54, 5862.CrossRefGoogle Scholar
Gu, H, Zou, YR, Rajewsky, K 1993. Independent control of immunoglobulin switch recombination at individual switch regions evidenced through Cre-loxP-mediated gene targeting. Cell 73, 11551164.CrossRefGoogle ScholarPubMed
Hammer, RE, Pursel, VG, Rexroad, CE Jr, Wall, RJ, Bolt, DJ, Ebert, KM, Palmiter, RD, Brinster, RL 1985. Production of transgenic rabbits, sheep and pigs by microinjection. Nature 315, 680683.CrossRefGoogle ScholarPubMed
Kühn, R, Schwenk, F, Aguet, M, Rajewsky, K 1995. Inducible gene targeting in mice. Science 269, 14271429.CrossRefGoogle ScholarPubMed
Lai, L, Kolber-Simonds, D, Park, KW, Cheong, HT, Greenstein, JL, Im, GS, Samuel, M, Bonk, A, Rieke, A, Day, BN, Murphy, CN, Carter, DB, Hawley, RJ, Prather, RS 2002. Production of alpha-1,3-galactosyltransferase knockout pigs by nuclear transfer cloning. Science 295, 10891092.CrossRefGoogle ScholarPubMed
Li, G, Robinson, GW, Lesche, R, Martinez-Diaz, H, Jiang, Z, Rozengurt, N, Wagner, KU, Wu, DC, Lane, TF, Liu, X, Hennighausen, L, Wu, H 2002. Conditional loss of PTEN leads to precocious development and neoplasia in the mammary gland. Development 129, 41594170.CrossRefGoogle ScholarPubMed
Miller, KF, Bolt, DJ, Pursel, VG, Hammer, RE, Pinkert, CA, Palmiter, RD, Brinster, RL 1989. Expression of human or bovine growth hormone gene with a mouse metallothionein-1 promoter in transgenic swine alters the secretion of porcine growth hormone and insulin-like growth factor-I. The Journal of Endocrinology 120, 481488.CrossRefGoogle ScholarPubMed
Raffai, RL, Weisgraber, KH 2002. Hypomorphic apolipoprotein E mice: a new model of conditional gene repair to examine apolipoprotein E-mediated metabolism. The Journal of Biological Chemistry 277, 1106411068.CrossRefGoogle ScholarPubMed
Sauer, B 1998. Inducible gene targeting in mice using the Cre/loxp system. Methods 14, 381392.CrossRefGoogle Scholar
Schmidt, EE, Taylor, DS, Prigge, JR, Barnett, S, Capecchi, MR 2000. Illegitimate Cre dependent chromosome rearrangements in transgenic mouse spermatics. Proceedings of the National Academy of Sciences of the United States of America 97, 1370213707.CrossRefGoogle Scholar
Schwenk, F, Kuhn, R, Angrand, PO, Rajewsky, K, Stewart, AF 1998. Temporally and spatially regulated somatic mutagenesis in mice. Nucleic Acids Research 26, 14271432.CrossRefGoogle ScholarPubMed
Thomas, AV, Palm, M, Broers, AD, Zezafoun, H, Desmecht, DJ 2006. Genomic structure, promoter analysis, and expression of the porcine (Sus scrofa) Mx1 gene. Immunogenetics 58, 383389.CrossRefGoogle ScholarPubMed
Tucker, A, Belcher, C, Moloo, B, Bell, J, Mazzulli, T, Humar, A, Hughes, A, McArdle, P, Talbot, A 2002. The production of transgenic pigs for potential use in clinical xenotransplantation: microbiological evaluation. Xenotransplantation 9, 191202.CrossRefGoogle ScholarPubMed