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Somatic cell nuclear transfer in zebrafish

Nature Methods volume 6, pages 733–735 (2009)Cite this article

Abstract

We developed a method for somatic cell nuclear transfer in zebrafish using laser-ablated metaphase II eggs as recipients, the micropyle for transfer of the nucleus and an egg activation protocol after nuclear reconstruction. We produced clones from cells of both embryonic and adult origins, although the latter did not give rise to live adult clones.

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Figure 1: Protocol for SCNT.
Figure 2: Phenotype of cloned zebrafish and its offspring.

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Acknowledgements

We thank K. Kawakami (National Institute of Genetics, Japan) for kindly donating transgenic TuLF zebrafish, Michigan Department of Natural Resources and egg takers at the little Manistee weir for facilitating CSOF collection; A. Busta, A. Mckay, S. Murthy, S. Helma and S. Fredline for maintaining the zebrafish facility; and the Royal Thai government and Kasetsart University for studentship of K.S. This project was supported by the College of Veterinary Medicine–endowed genetic research fund, the Michigan State University Experiment Station, the office of the Vice President for Research and Graduate Studies, the Michigan State University foundation, and the Naylor Family Foundation.

Author information

Authors and Affiliations

  1. Comparative Medicine and Integrative Biology, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan, USA

    Kannika Siripattarapravat

  2. Department of Animal Science, Michigan State University, East Lansing, Michigan, USA

    Boonya Pinmee & Jose B Cibelli

  3. Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, Michigan, USA

    Patrick J Venta

  4. Department of Small Animal Clinical Sciences, Michigan State University, East Lansing, Michigan, USA

    Patrick J Venta

  5. Department of Pediatrics/Human Development, Michigan State University, East Lansing, Michigan, USA

    Chia-Cheng Chang

  6. Department of Physiology, Michigan State University, East Lansing, Michigan, USA

    Jose B Cibelli

  7. Programa Andaluz de Terapia Celular y Medicina Regenerativa, Andalucía, Spain

    Jose B Cibelli

Authors
  1. Kannika Siripattarapravat
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  2. Boonya Pinmee
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  3. Patrick J Venta
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  4. Chia-Cheng Chang
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  5. Jose B Cibelli
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Contributions

J.B.C. initiated and supervised the whole project. K.S. and J.B.C. conceived the cloning methodology, designed the experiments, analyzed data, and wrote the manuscript. K.S. and B.P. performed the SCNT experiment. The SNP genotyping was designed by P.J.V. and K.S. and performed by K.S. The cell culture protocol was optimized by K.S. under supervision of C.-C.C. The zebrafish facility was maintained by B.P. and K.S.

Corresponding author

Correspondence to Jose B Cibelli.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–3 and Supplementary Tables 1–2 (PDF 487 kb)

Supplementary Movie 1

Zebrafish SCNT movie. (MOV 54235 kb)

Supplementary Movie 2

Clones and their offspring. (AVI 12102 kb)

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Siripattarapravat, K., Pinmee, B., Venta, P. et al. Somatic cell nuclear transfer in zebrafish. Nat Methods 6, 733–735 (2009). https://doi.org/10.1038/nmeth.1369

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  • DOI: https://doi.org/10.1038/nmeth.1369

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