Transgene expression in Xenopus rods.
The photoreceptors of the vertebrate retina express a large number of proteins that are involved in the process of light transduction. These genes appear to be coordinately regulated at the level of transcription, with rod- and cone-specific isoforms (J. Hurley (1992) J. Bioenerg. Biomembr. 24, 219-226). The mechanisms that regulate gene expression in a rod/cone-specific fashion have been difficult to address using traditional approaches and remain unknown. Regulation of the phototransduction proteins is medically important, since mutations in several of them cause retinal degeneration (P. Rosenfeld and T. Dryja (1995) in: Molecular Genetics of Ocular Disease (J.L. Wiggs, Ed.), pp. 99-126, Wiley-Liss Inc.). An experimental system for rapidly producing retinas expressing a desired mutant would greatly facilitate investigations of retinal degeneration. We report here that transgenic frog embryos (K. Kroll and E. Amaya (1996) Development 122, 3173-3183) can be used to study cell-specific expression in the retina. We have used a 5.5 kb 5' upstream fragment from the Xenopus principal rod opsin gene (S. Batni et al. (1996) J. Biol. Chem. 271, 3179-3186) controlling a reporter gene, green fluorescent protein (GFP), to produce numerous independent transgenic Xenopus. We find that this construct drives expression only in the retina and pineal, which is apparent by 4 days post-nuclear injection. These are the first results using transgenic Xenopus for retinal promoter analysis and the potential for the expression in rod photoreceptors of proteins with dominant phenotypes.
Knox
B E
BE
Department of Biochemistry and Molecular Biology, SUNY Health Science Center at Syracuse, NY 13210, USA. knoxb@hscsyr.edu
Schlueter
C
C
Sanger
B M
BM
Green
C B
CB
Besharse
J C
JC
D000818:Animals; D030801:Animals, Genetically Modified; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012160:Retina / Q000378:metabolism*; D012243:Rhodopsin / Q000235:genetics*; D014982:Xenopus laevis
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D012243:Rhodopsin