Transgenic approaches to retinal development and function in Xenopus laevis.
The African clawed frog Xenopus laevis has long been used to study the development and function of the vertebrate retina. An efficient technique for generating transgenic Xenopus embryos, the REMI procedure, has enabled the stable overexpression of transgenes in developing and mature X. laevis. In the retina, transgenes driven by retinal-specific promoters have been used to study protein trafficking, circadian rhythms, and retinal degeneration. The REMI technique is surprisingly simple, consisting of integration of plasmid DNA into permeabilized sperm nuclei, followed by transplantation of these nuclei into unfertilized eggs. Here, we describe the reagents and steps necessary for generation of transgenic embryos using the REMI reaction and discuss its applications for the study of retinal development.
Hutcheson
David A
DA
Department of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City 84132, USA.
Vetter
Monica L
ML
D000818:Animals; D030801:Animals, Genetically Modified; D002467:Cell Nucleus / Q000502:physiology; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008297:Male; D023421:Models, Animal; D010063:Ovum / Q000502:physiology; D012160:Retina / Q000254:growth & development* / Q000502:physiology*; D013094:Spermatozoa / Q000502:physiology; D019076:Transgenes / Q000502:physiology*; D014982:Xenopus laevis / Q000235:genetics*