Generation of transgenic Xenopus using restriction enzyme-mediated integration

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

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified*
  • Cell Nucleus
  • Cell Separation
  • DNA Restriction Enzymes / metabolism*
  • Female
  • Male
  • Nuclear Transfer Techniques
  • Ovum / cytology
  • Spermatozoa / cytology
  • Xenopus laevis / genetics*

Substances

  • DNA Restriction Enzymes