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Somatic cell nuclear transfer in zebrafish

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.

References

  1. Anderson, K.V. & Ingham, P.W. Nat. Genet. 33, 285–293 (2003).

    Article  CAS  Google Scholar 

  2. Fan, L., Moon, J., Crodian, J. & Collodi, P. Transgenic Res. 15, 21–30 (2006).

    Article  CAS  Google Scholar 

  3. Ma, C. et al. Proc. Natl. Acad. Sci. USA 98, 2461–2466 (2001).

    Article  CAS  Google Scholar 

  4. Lee, K.Y. et al. Nat. Biotechnol. 20, 795–799 (2002).

    Article  CAS  Google Scholar 

  5. Siripattarapravat, K., Busta, A., Steibel, J.P. & Cibelli, J. Zebrafish 6, 97–105 (2009).

    Article  CAS  Google Scholar 

  6. Egli, D., Rosains, J., Birkhoff, G. & Eggan, K. Nature 447, 679–685 (2007).

    Article  CAS  Google Scholar 

  7. Lamason, R.L. et al. Science 310, 1782–1786 (2005).

    Article  CAS  Google Scholar 

  8. Nagayoshi, S. et al. Development 135, 159–169 (2008).

    Article  CAS  Google Scholar 

  9. Pauls, S., Geldmacher-Voss, B. & Campos-Ortega, J.A. Dev. Genes Evol. 211, 603–610 (2001).

    Article  CAS  Google Scholar 

  10. Amores, A. & Postlethwait, J.H. Methods Cell Biol. 60, 323–338 (1999).

    Article  CAS  Google Scholar 

  11. Westerfield, M. The Zebrafish Book, 4 ed. (University of Oregon Press, Eugene, Oregon, USA, 2000).

    Google Scholar 

  12. Lin, T.M. et al. Stem Cells Dev. 14, 92–102 (2005).

    Article  CAS  Google Scholar 

  13. Kent, W.J. Genome Res. 12, 656–664 (2002).

    Article  CAS  Google Scholar 

  14. Rozen, S. & Skaletsky, H. Methods Mol. Biol. 132, 365–386 (2000).

    CAS  Google Scholar 

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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

Authors

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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