Generation of transgenic Xenopus using restriction enzyme-mediated integration.
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.
Department of Neuroscience & Physiology, SUNY Upstate Medical University, Syracuse, NY, USA.
Mohammad
M
Haeri
Barry E
BE
Knox
D000818:Animals; D030801:Animals, Genetically Modified; D002467:Cell Nucleus; D002469:Cell Separation; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D005260:Female; D008297:Male; D053652:Nuclear Transfer Techniques; D010063:Ovum / Q000166:cytology; D013094:Spermatozoa / Q000166:cytology; D014982:Xenopus laevis / Q000235:genetics*
D004262:DNA Restriction Enzymes