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Development of Cre–loxP technology in zebrafish to study the regulation of fish reproduction

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Abstract

One cannot seek permission to market transgenic fish mainly because there is no field test or any basic research on technological developments for evaluating their biosafety. Infertility is a necessary adjunct to exploiting transgenic fish unless completely secure land-locked facilities are available. In this study, we report the generation of a Cre transgenic zebrafish line using a cytomegalovirus promoter. We also produced fish carrying the Bax1 and Bax2 plasmids; these genes were separated by two loxP sites under a zona pellucida C promoter or were driven by an anti-Müllerian hormone promoter. We inserted a red fluorescent protein gene between the two loxP sites. After obtaining transgenic lines with the two transgenic fish crossed with each other (Cre transgenic zebrafish x loxP transgenic zebrafish), the floxed DNA was found to be specifically eliminated from the female or male zebrafish, and apoptosis gene expressions caused ovarian and testicular growth cessation and degeneration. Overexpression of the Bax1 and Bax2 genes caused various expression levels of apoptosis-related genes. Accordingly, this transgenic zebrafish model system provides a method to produce infertile fish and may be useful for application to genetically modified fish.

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Acknowledgments

This work was supported by a grant from the Development Program of Industrialization for Agricultural Biotechnology (money provided by Academia Sinica), called “Study and development of novel pattern of transgenic fluorescent ornamental fish and infertility technology” to Dr. Jyh-Yih Chen in 2010 and 2011. The Cre–loxP plasmid was kindly provided as a gift from Dr. Allan Bradley (Wellcome Trust Sanger Institute, Cambridge, UK).

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Authors and Affiliations

  1. Graduate Institute of Engineering Technology-Doctoral, National Taipei University of Technology, 1 Chung-Hsiao E. Rd., Sec. 3, Taipei, 10608, Taiwan

    Heng-Ju Lin & Yeh-Fang Duann

  2. Marine Research Station, Institute of Cellular and Organismic Biology, Academia Sinica, 23-10 Dahuen Rd., Jiaushi, Ilan, 262, Taiwan

    Shu-Hua Lee & Jyh-Yih Chen

  3. Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan

    Jen-Leih Wu

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  1. Heng-Ju Lin
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  2. Shu-Hua Lee
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  3. Jen-Leih Wu
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  4. Yeh-Fang Duann
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  5. Jyh-Yih Chen
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Correspondence to Jyh-Yih Chen.

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Lin, HJ., Lee, SH., Wu, JL. et al. Development of Cre–loxP technology in zebrafish to study the regulation of fish reproduction. Fish Physiol Biochem 39, 1525–1539 (2013). https://doi.org/10.1007/s10695-013-9806-6

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  • DOI: https://doi.org/10.1007/s10695-013-9806-6

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