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Transgenic Zebrafish

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 136))

Abstract

The zebrafish has become a popular model system for genetic studies of vertebrate development. Zebrafish are amenable to such studies because the generation time of zebrafish is short, adults are small and easy to maintain, and females typically lay up to several hundred eggs per mating. Early development occurs rapidly outside the mother and the embryos are transparent, making the observation and manipulation of the embryos quite simple (1). Recently, large-scale chemical mutagenesis screens in zebrafish have identified over 1000 mutations, including hundreds of genes essential for embryogenesis (2,3). However, to maximally benefit from genetic analysis of embryogenesis using zebrafish, a number of additional challenges must be overcome. First, the chemically induced mutant genes must be identified. Second, the mutations must be analyzed in vivo at the cellular and molecular levels. Third, those functional genes that have been missed by the two screens must be identified. The generation of transgenic zebrafish can provide useful tools for overcoming each of these challenges.

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References

  1. Kimmel, C. B. (1989) Genetics and early development of zebrafish. Trends Genet. 8, 283–288.

    Article  Google Scholar 

  2. Driever, W., Solnica-Krezel, L., Schier, A. F., Neuhauss, S. C., Malicki, J., Stemple, D. L., Stainier, D. Y., Zwartkruis, F., Abdelilah, S., Rangini, Z., Belak, J., and Boggs, C. (1996) A genetic screen for mutations affecting embryogenesis in zebrafish. Development 123, 37–46.

    PubMed  CAS  Google Scholar 

  3. Haffter, P., Granato, M., Brand, M., Mullins, M. C., Hammerschmidt, M., Kane, D. A., Odenthal, J., van Eeden, F. J., Jiang, Y. J., Heisenberg, C. P., Kelsh, R. N., Furutani-Seiki, M., Vogelsang, E., Beuchle, D., Schach, U., Fabian, C., and Nusslein-Volhard, C. (1996) The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio. Development 123, 1–36.

    PubMed  CAS  Google Scholar 

  4. Stuart, G. W., McMurray, J. V., and Westerfield, M. (1988) Replication, integration and stable germ-line transmission of foreign sequences injected into early zebrafish embryos. Development 103, 403–12.

    PubMed  CAS  Google Scholar 

  5. Stuart, G. W., Vielkind, J. R., McMurray, J. V., and Westerfield, M. (1990) Stable lines of transgenic zebrafish exhibit reproducible patterns of transgene expression. Development 109, 577–584.

    PubMed  CAS  Google Scholar 

  6. Amsterdam, A., Lin, S., and Hopkins, N. (1995) The Aequorea victoria green fluorescent protein can be used as a reporter in live zebrafish embryos. Dev. Biol. 171, 123–129.

    Article  PubMed  CAS  Google Scholar 

  7. Bayer, T. A., and Campos-Ortega, J. A. (1992) A transgene containing lacZ is expressed in primary sensory neurons in zebrafish. Development 115, 421–426.

    PubMed  CAS  Google Scholar 

  8. Lin, S., Yang, S., and Hopkins, N. (1994b) lacZ expression in germline transgenic zebrafish can be detected in living embryos. Dev. Biol. 161, 77–83.

    Article  PubMed  Google Scholar 

  9. Long, Q., Meng, A., Wang, H., Jessen, J. R., Farrell, M. J., and Lin, S. (1997) GATA-1 expression pattern can be recapitulated in living transgenic zebrafish using GFP reporter gene. Development 124, 4105–4111.

    PubMed  CAS  Google Scholar 

  10. Culp, P., Nusslein-Volhard, C., and Hopkins, N. (1991) High-frequency germ-line transmission of plasmid DNA sequences injected into fertilized zebrafish eggs. Proc. Natl. Acad. Sci. USA 88, 7953–7957.

    Article  PubMed  CAS  Google Scholar 

  11. Gibbs, P. D., Gray, A., and Thorgaard, G. (1994) Inheritance of P element and reporter gene sequences in zebrafish. Mol. Marine Biol. Biotechnol. 3, 317–326.

    PubMed  CAS  Google Scholar 

  12. Lin, S., Gaiano, N., Culp, P., Burns, J. C., Friedmann, T., Yee, J. K., and Hopkins, N. (1994a) Integration and germ-line transmission of a pseudotyped retroviral vector in zebrafish. Science 265, 666–669.

    Article  PubMed  CAS  Google Scholar 

  13. Zelenin, A. V., Alimov, A. A., Barmintzev, V. A., Beniumov, A. O., Zelenina, I. A., Krasnov, A. M., and Kolesnikov, V. A. (1991) The delivery of foreign genes into fertilized fish eggs using high-velocity microprojectiles. FEBS Lett. 287, 118–120.

    Article  PubMed  CAS  Google Scholar 

  14. Muller, F., Lele, Z., Varadi, L., Menczel, L., and Orban, L. (1993) Efficient transient expression system based on square pulse electroporation and in vivo luciferase assay of fertilized fish eggs. FEBS Lett. 324, 27–32.

    Article  PubMed  CAS  Google Scholar 

  15. Powers, D. A., Hereford, L., Cole, T., Chen, T. T., Lin, C. M., Kight, K., Creech, K., and Dunham, R. (1992) Electroporation: a method for transferring genes into the gametes of zebrafish (Brachydanio rerio), channel catfish (Ictalurus punctatus), and common carp (Cyprinus carpio) Mol. Marine Biol. Biotechnol. 1, 301–308.

    PubMed  CAS  Google Scholar 

  16. Meng, A., Tang, H., Ong, B. A., Farrell, M. J., and Lin, S. (1997) Promoter analysis in living zebrafish embryos identifies a cis-acting motif required for neuronal expression of GATA-2. Proc. Natl. Acad. Sci. USA 94, 6267–6272.

    Article  PubMed  CAS  Google Scholar 

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© 2000 Humana Press Inc.

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Lin, S. (2000). Transgenic Zebrafish. In: Tuan, R.S., Lo, C.W. (eds) Developmental Biology Protocols: Volume II. Methods in Molecular Biology™, vol 136. Humana Press. https://doi.org/10.1385/1-59259-065-9:375

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  • DOI: https://doi.org/10.1385/1-59259-065-9:375

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-853-0

  • Online ISBN: 978-1-59259-065-0

  • eBook Packages: Springer Protocols

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