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Methods Mol Biol. 2012;884:17-39. doi: 10.1007/978-1-61779-848-1_2.

Generation of transgenic Xenopus using restriction enzyme-mediated integration.

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

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.

PMID:
22688696
DOI:
10.1007/978-1-61779-848-1_2
[PubMed - indexed for MEDLINE]

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