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Pig transgenesis by piggyBac transposition in combination with somatic cell nuclear transfer

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Abstract

The production of animals by somatic cell nuclear transfer (SCNT) is inefficient, with approximately 2 % of micromanipulated oocytes going to term and resulting in live births. However, it is the most commonly used method for the generation of cloned transgenic livestock as it facilitates the attainment of transgenic animals once the nuclear donor cells are stably transfected and more importantly as alternatives methods of transgenesis in farm animals have proven even less efficient. Here we describe piggyBac-mediated transposition of a transgene into porcine primary cells and use of these genetically modified cells as nuclear donors for the generation of transgenic pigs by SCNT. Gene transfer by piggyBac transposition serves to provide an alternative approach for the transfection of nuclear donor cells used in SCNT.

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Acknowledgments

This study was supported by a grant from the National Science Foundation for Young Scholars of China (Grant Number: 31101689), a grant from the National High Technology Research and Development Program of China (863 Program, Grant Number: 2011AA100304), a grant from Department of Science and Technology of Guangdong, China (Grant Number: 2011A020901001) and by National Institutes of Health Grants 5P20RR024206 and R01 GM083158-01A1 (to S.M.).

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Authors and Affiliations

  1. Department of Animal Genetics, Breeding and Reproduction, South China Agricultural University, The New Building of College of Animal Science, Room 315, Guangzhou, 510642, Guangdong, People’s Republic of China

    Zhenfang Wu, Zhiqian Xu, Xian Zou, Fang Zeng & Zicong Li

  2. Department of Animal Production, College of Animal Science, South China Agricultural University, Guangzhou, 510642, Guangdong, People’s Republic of China

    Dewu Liu

  3. Wen’s Research Institute, Guangdong Wen’s Food Group Co. Ltd., Yunfu, 527439, Guangdong, People’s Republic of China

    Junsong Shi

  4. Department of Anatomy, Biochemistry and Physiology, Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii at Manoa, E-124, 1960 East–West Rd., Honolulu, HI, 96822, USA

    Johann Urschitz & Stefan Moisyadi

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  1. Zhenfang Wu
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  2. Zhiqian Xu
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  3. Xian Zou
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  4. Fang Zeng
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  5. Junsong Shi
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  6. Dewu Liu
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  7. Johann Urschitz
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  8. Stefan Moisyadi
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  9. Zicong Li
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Corresponding authors

Correspondence to Stefan Moisyadi or Zicong Li.

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Wu, Z., Xu, Z., Zou, X. et al. Pig transgenesis by piggyBac transposition in combination with somatic cell nuclear transfer. Transgenic Res 22, 1107–1118 (2013). https://doi.org/10.1007/s11248-013-9729-0

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  • DOI: https://doi.org/10.1007/s11248-013-9729-0

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