Hostname: page-component-586b7cd67f-dlnhk Total loading time: 0 Render date: 2024-11-26T14:00:58.642Z Has data issue: false hasContentIssue false

Recloned transgenic pigs possess normal reproductive performance and stable genetic transmission capacity

Published online by Cambridge University Press:  12 July 2012

Zubing Cao*
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China.
Yan Li
Affiliation:
Beijing Gene & Protein Biotechnology Co., Ltd., Beijing 100193, China.
Xiao Wen
Affiliation:
Beijing Gene & Protein Biotechnology Co., Ltd., Beijing 100193, China.
Zhiyuan Li
Affiliation:
Beijing Gene & Protein Biotechnology Co., Ltd., Beijing 100193, China.
Changsheng Mi
Affiliation:
Beijing Gene & Protein Biotechnology Co., Ltd., Beijing 100193, China.
Zaihu Zhang
Affiliation:
Beijing Gene & Protein Biotechnology Co., Ltd., Beijing 100193, China.
Ning Li
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China.
Qiuyan Li*
Affiliation:
State Key Laboratory for Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China.
*
All correspondence to: Zubing Cao. State Key Laboratory for Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China. Tel:/Fax: +86 10 62731142. e-mail: [email protected]
Qiuyan Li, State Key Laboratory for Agrobiotechnology, College of Biological Science, China Agricultural University, Beijing 100193, China. Tel:/Fax: +86 10 62731143. e-mail: [email protected]

Summary

The present study investigated whether a recloning procedure would affect the reproductive performance or the germline transmission capacity of recloned transgenic pigs. This study has also laid the foundation for the development of elite transgenic swine breeds in the future. Recloned transgenic pigs were developed from ear tissue fibroblasts of primary transgenic cloned pigs using a recloning procedure, and their reproductive performance and exogenous gene transmission were analyzed. Two transgenic cell lines with different genetic backgrounds (derived from a female miniature pig and a male Landrace pig) with stable expression of green fluorescent protein (GFP) were established successfully. Furthermore, recloned transgenic embryos were developed to full term successfully. One female Chinese experimental miniature piglet (CEMP) (GFP+) and three male Landrace piglets (GFP+) were delivered naturally. Furthermore, the index values for the reproductive characteristics of the recloned transgenic pigs, such as puberty, gestation period, sperm volume and sperm concentration, were not significantly different from those of conventionally bred pigs. In addition, 53% of the F1 offspring of the recloned transgenic pigs were GFP positive. These results demonstrate that ear tissue fibroblasts from primary transgenic cloned pigs efficiently support the full-term development of recloned transgenic embryos. Furthermore, recloned transgenic pigs maintain normal reproductive performance and stable germline (genetic) transmission capacities.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2012 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Ahn, K.S., Kim, Y.J., Kim, M., Lee, B.H., Heo, S.Y., Kang, M.J., Kang, Y.K., Lee, J.W., Lee, K.K., Kim, J.H., Nho, W.G., Hwang, S.S., Woo, J.S., Park, J.K., Park, S.B. & Shim, H. (2011). Resurrection of an alpha-1,3-galactosyltransferase gene-targeted miniature pig by recloning using postmortem ear skin fibroblasts. Theriogenology 75, 933–9.Google Scholar
Cho, S.J., Bang, J.I., Yu, X.F., Lee, Y.S., Kim, J.H., Jeon, J.T., Yee, S.T. & Kong, I.K. (2010). Generation of a recloned transgenic cat expressing red fluorescence protein. Theriogenology 73, 848–55.Google Scholar
Cho, S.K., Kim, J.H., Park, J.Y., Choi, Y.J., Bang, J.I., Hwang, K.C., Cho, E.J., Sohn, S.H., Uhm, S.J., Koo, D.B., Lee, K.K., Kim, T. & Kim, J.H. (2007). Serial cloning of pigs by somatic cell nuclear transfer: restoration of phenotypic normality during serial cloning. Dev. Dynam. 236, 3369–82.Google Scholar
Fujimura, T., Murakami, H., Kurome, M., Takahagi, Y., Shigehisa, T. & Nagashima, H. (2008). Effects of recloning on the efficiency of production of α1,3-galactosyltransferase knockout pigs. J. Reprod. Dev. 54, 5862.Google Scholar
Hill, J.R., Winger, Q.A., Long, C.R., Looney, C.R., Thompson, J.A. & Westhusin, M.E. (2000). Development rates of male bovine nuclear transfer embryos derived from adult and fetal cells. Biol. Reprod. 62, 1135–40.Google Scholar
Hill, J.R., Winger, Q.A., Burghardt, R.C. & Westhusin, M.E. (2001). Bovine nuclear transfer embryo development using cells derived from a cloned fetus. Anim. Reprod. Sci. 67, 1726.Google Scholar
Kuroiwa, Y., Kasinathan, P., Sathiyaseelan, T., Jiao, J.A., Matsushita, H., Sathiyaseelan, J., Wu, H., Mellquist, J., Hammitt, M., Koster, J., Kamoda, S., Tachibana, K., Ishida, I. & Robl, J.M. (2009). Antigen-specific human polyclonal antibodies from hyperimmunized cattle. Nat. Biotechnol. 27, 173–81.Google Scholar
Kurome, M., Hilsatomi, H., Matsumoto, S., Tomii, R., Ueno, S., Hiruma, K., Saito, H., Nakamura, K., Okumura, K., Matsumoto, M., Kaji, Y., Endo, F. & Nagashima, H. (2008). Production efficiency and telomere length of the cloned pigs following serial somatic cell nuclear transfer. J. Reprod. Dev. 54, 254–8.CrossRefGoogle ScholarPubMed
Li, R.F., Lai, L.X., Wax, D., Hao, Y.H., Murphy, C.N., Rieke, A., Samuel, M., Linville, M.L., Korte, S.W., Evans, R.W., Turk, J.R., Kang, J.X., Witt, W.T., Dai, Y.F. & Prather, R.S. (2006). Cloned transgenic swine via in vitro production and cryopreservation. Biol. Reprod. 75, 226–30.Google Scholar
Liu, H.B., Lv, P.R., He, R.G., Yang, X.G., Qin, X.E., Pan, T.B., Huang, G.Y., Huang, M.R., Lu, Y.Q., Lu, S.S., Li, D.S. & Lu, K.H. (2010). Cloned guangxi bama minipig (Sus scrofa) and its offspring have normal reproductive performance. Cell. Reprog. 12, 543–50.Google Scholar
Martin, M., Adams, C. & Wiseman, B. (2004). Pre-weaning performance and health of pigs born to cloned (fetal cell derived) swine versus non-cloned swine. Theriogenology 62, 113–22.Google Scholar
Nagashima, H., Kashiwazaki, N., Ashman, R.J., Grupen, C.G. & Nottle, M.B. (1995). Cryopreservation of porcine embryos. Nature 374, 416.CrossRefGoogle ScholarPubMed
Park, K.W., Lai, L.X., Cheong, H.T., Cabot, R., Sun, Q.Y., Wu, G.M., Rucker, E.B., Durtschi, D., Bonk, A., Samuel, M., Rieke, A., Day, B.N., Murphy, C.N., Carter, D.B. & Prather, R.S. (2002). Mosaic gene expression in nuclear transfer-derived embryos and the production of cloned transgenic pigs from ear-derived fibroblasts. Biol. Reprod. 66, 1001–5.Google Scholar
Park, K.W., Choi, K.M., Hong, S.P., Han, G.S., Yoo, J.Y., Jin, D.I., Seol, J.G. & Park, C.S. (2008). Production of transgenic recloned piglets harboring the human granulocyte–macrophage colony stimulating factor (hGM-CSF) gene from porcine fetal fibroblasts by nuclear transfer. Theriogenology 70, 1431–8.Google Scholar
Shibata, M., Otake, M., Tsuchiya, S., Chikyu, M., Horiuchi, A., & Kawarasaki, T. (2006). Reproductive and growth performance in jin hua pigs cloned from somatic cell nuclei and the meat quality of their offspring. J. Reprod. Dev. 52, 583–90.Google Scholar
Uhm, S.J., Gupta, M.K., Das, Z.C., Kim, J.H., Park, C., Kim, T. & Lee, H.T. (2009). Effect of transgene introduction and recloning on efficiency of porcine transgenic cloned embryo production in vitro. Reprod. Domes. Anim. 44, 106–15.Google Scholar
Wakai, T., Tanaka, H., Yamanaka, K., Sugimura, S., Sasada, H., Kawahara, M., Kobayashi, E. & Sato, E. (2008). Induction of estrus in pubertal miniature gilts. Anim. Reprod. Sci. 103, 193–8.Google Scholar
Wang, S., Sun, X., Ding, F., Zhang, K., Zhao, R., Li, S., Li, R., Tang, B., Zhang, L., Liu, Y., Li, J., Gao, F., Wang, H., Wang, L., Dai, Y. & Li, N. (2009). Removal of selectable marker gene from fibroblast cells in transgenic cloned cattle by transient expression of Cre recombinase and subsequent effects on recloned embryo development. Theriogenology 72, 535–41.Google Scholar
Yang, X.Z., Smith, S.L., Tian, X.C., Lewin, H.A., Renard, J.P. & Wakayama, T. (2007). Nuclear reprogramming of cloned embryos and its implications for therapeutic cloning. Nat. Genet. 39, 295302.Google Scholar
Yeste, M., Barrera, X., Coll, D. & Bonet, S. (2011). The effects on boar sperm quality of dietary supplementation with omega-3 polyunsaturated fatty acids differ among porcine breeds. Theriogenology 76, 184–96.Google Scholar
Yoshioka, K., Suzuki, C., Tanaka, A., Anas, I.M.K. & Iwamura, S. (2002). Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol. Reprod. 66, 112–9.Google Scholar
Zhang, Y.H., Li, J., Villemoes, K., Pedersen, A.M., Purup, S. & Vajta, G. (2007). An epigenetic modifier results in improved in vitro blastocyst production after somatic cell nuclear transfer. Cloning Stem Cells 9, 357–63.Google Scholar