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Cloned zebrafish by nuclear transfer from long-term-cultured cells

Nature Biotechnology volume 20, pages 795–799 (2002)Cite this article

Abstract

Although mammals have been cloned from genetically manipulated cultured cells, a comparable achievement has not been realized in lower vertebrates. Here we report that fertile transgenic zebrafish can be obtained by nuclear transfer using embryonic fibroblast cells from long-term cultures. The donor nuclei, modified by retroviral insertions expressing green fluorescent protein (GFP), were transplanted into manually enucleated eggs. Overall, a 2% success rate was achieved, resulting in 11 adult transgenic zebrafish expressing GFP. These nuclear transplants produced fertile, diploid offspring, and their F1/F2 progeny continued to express GFP in a pattern identical to that of the founder fish. This finding demonstrates that slowly dividing nuclei from cultured cells can be reprogrammed to support rapid embryonic development and sets up a foundation for targeted genetic manipulation in zebrafish.

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Figure 1: Zebrafish recipient eggs used for nuclear transfer.
Figure 2: Enucleation and nuclear transfer in zebrafish.
Figure 3: Characterization of cultured embryonic fibroblast cells.
Figure 4: Transgenic zebrafish embryos produced by nuclear transfer.

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Acknowledgements

We thank members of our laboratory for discussion and technical assistance. We thank J. Chen for critically reading the manuscript. This work was supported by a grant from the National Institutes of Health (R01 RR13227) to S.L.

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Authors and Affiliations

  1. Department of Molecular, Cellular, and Developmental Biology, University of California Los Angeles, Los Angeles, 90095-1606, CA

    Ki-Young Lee, Haigen Huang, Bensheng Ju, Zhongan Yang & Shuo Lin

  2. School of Agricultural Biotechnology, Seoul National University, Seoul, 151-742, Korea

    Ki-Young Lee

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Correspondence to Shuo Lin.

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Lee, KY., Huang, H., Ju, B. et al. Cloned zebrafish by nuclear transfer from long-term-cultured cells. Nat Biotechnol 20, 795–799 (2002). https://doi.org/10.1038/nbt721

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