Transgenic Xenopus laevis tadpoles: a transient in vivo model system for the manipulation of lens function and lens development.
Rodent gamma-crystallin promoters were recognized as lens-specific promoters in micro-injected Xenopus laevis tadpoles and targeted the expression of the chloramphenicol acetyl transferase (CAT) reporter gene to the tadpole lens. The onset of expression coincided with lens cell formation. The level of expression continued to increase up to 9 days of development (stage 47), stayed at that level till at least day 13 and dropped by only 57% at day 21. In contrast, the level of expression of a non-tissue-specific promoter, the SV40 early promoter, decreased rapidly in the eye during development and was only detectable up to stage 44 (day 5). The stability of the CAT activity in the lens was assessed by delivering a pulse of activity from a heat shock promoter-CAT fusion gene. The half-life of the CAT activity in the eye was the same as that in the tail. The increase in CAT activity in the lens thus depends upon continued activity of the injected gamma-crystallin promoters. Our data demonstrate that mammalian promoters can be used to target gene expression to specific tissues during Xenopus laevis development.
Brakenhoff
R H
RH
Laboratory of Molecular Biology, University of Nijmegen, The Netherlands.
Ruuls
R C
RC
Jacobs
E H
EH
Schoenmakers
J G
JG
Lubsen
N H
NH
D000818:Animals; D030801:Animals, Genetically Modified; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D003459:Crystallins / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D007908:Lens, Crystalline / Q000201:enzymology / Q000254:growth & development / Q000502:physiology*; D051379:Mice; D008845:Microinjections; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D016385:TATA Box; D014982:Xenopus laevis
D003459:Crystallins; D015500:Chloramphenicol O-Acetyltransferase