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Generation of Transgenic Xenopus Using Restriction Enzyme-Mediated Integration

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Retinal Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 884))

Abstract

Transgenesis, the process of incorporating an exogenous gene (transgene) into an organism’s genome, is a widely used tool to develop models of human diseases and to study the function and/or regulation of genes. Generating transgenic Xenopus is rapid and involves simple in vitro manipulations, taking advantage of the large size of the amphibian egg and external embryonic development. Restriction enzyme-mediated integration (REMI) has a number of advantages for transgenesis compared to other methods used to produce transgenic Xenopus, including relative efficiency, higher transgene expression levels, fewer genetic chimera in founder transgenic animals, and near-complete germ-line transgene transmission. This chapter explains the REMI method for generating transgenic Xenopus laevis tadpoles, including improvements developed to enable studies in the mature retina.

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Acknowledgments

We thank Annabelle Pellerin and Maria Goralski for help in preparation of this manuscript. We acknowledge present and former lab and Center for Vision Research members who have helped develop this procedure over many years. This work was supported by the National Institutes of Health Grants EY-11256 and EY-12975 (B.E.K.), Research to Prevent Blindness (Unrestricted Grant to SUNY UMU Department of Ophthalmology), Fight for Sight (FFS) and Lions of CNY.

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Authors and Affiliations

  1. Department of Neuroscience & Physiology, SUNY Upstate Medical University, Syracuse, NY, USA

    Mohammad Haeri & Barry E. Knox

Authors
  1. Mohammad Haeri
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  2. Barry E. Knox
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Corresponding author

Correspondence to Barry E. Knox .

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Editors and Affiliations

  1. University of Alabama at Birmingham, University Blvd 444B, 1670, Birmingham, 35294-0019, Alabama, USA

    Shu-Zhen Wang

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Haeri, M., Knox, B.E. (2012). Generation of Transgenic Xenopus Using Restriction Enzyme-Mediated Integration. In: Wang, SZ. (eds) Retinal Development. Methods in Molecular Biology, vol 884. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-848-1_2

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  • DOI: https://doi.org/10.1007/978-1-61779-848-1_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-847-4

  • Online ISBN: 978-1-61779-848-1

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