In utero complementation of a neural crest-derived melanocyte defect using cell directed gene transfer.
This study describes an in utero approach for overexpressing genes in a cell-type directed manner. It uses an avian leukosis retroviral expression system coupled with a transgenic mouse line expressing the viral receptor tv-a from a tissue-specific promoter (RCAS-TVA system) (Federspiel et al., 1994, and reviewed in Fisher et al., 1999). A transgenic mouse line was generated expressing tv-a from the Dopachrome tautomerase promoter (DCT-tv-a) in embryonic melanocyte precursors (melanoblasts). RCAS virus encoding beta-galactosidase (RCAS-LacZ) or tyrosinase (RCAS-Tyr) was injected in utero into embryonic day 12.5 albino (tyrosinase inactive) mouse embryos. Animals were analyzed for beta-galactosidase activity or tyrosinase activity (hair pigmentation). RCAS gene expression was detected in 44% and 25% of the transgenic mice, respectively. We demonstrate the RCAS-TVA system coupled with the DCT-tv-a line of mice can be used for in utero infection.
Dunn
K J
KJ
Mouse Embryology Section, Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892-4472, USA.
Incao
A
A
Watkins-Chow
D
D
Li
Y
Y
Pavan
W J
WJ
D000818:Animals; D001354:Avian Leukosis Virus / Q000235:genetics / Q000302:isolation & purification; D030161:Avian Proteins; D002454:Cell Differentiation; D004622:Embryo, Mammalian / Q000166:cytology; D018014:Gene Transfer Techniques; D005816:Genetic Complementation Test; D008544:Melanocytes / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics; D011991:Receptors, Virus / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D014162:Transfection; D051153:Wnt Proteins; D029961:Zebrafish Proteins
D030161:Avian Proteins; D011518:Proto-Oncogene Proteins; D011991:Receptors, Virus; D011994:Recombinant Proteins; C088745:Tva receptor; D051153:Wnt Proteins; D029961:Zebrafish Proteins