International Mammalian Genome Society

The 16th International Mouse Genome Conference (2002)


Oral Presentation

Monday 18 November

17:15 - 17:30 HRS

COMPLEMENTATION OF MOUSE NEURAL CREST DEFECTS USING LINEAGE DIRECTED GENE TRANSFER

WJ Pavan
National Human Genome Research Institute

Co-Authors: Dunn KJ, Hou L, Loftus SK.
Institutions: Genetics Disease Research Branch, National Human Genome Research Institute, National Institutes of Health

Ectopic gene expression in fetal mouse tissues has been a powerful tool for analyzing gene function during development and testing for functional complementation of mouse mutant loci.  We are using retroviral infection to overexpress genes in a lineage directed manner in mouse embryos in vivo and tissue explants in vitro.  The approach utilizes an avian leukosis retroviral expression system coupled with a transgenic mouse line expressing the viral receptor tv-a  from a tissue specific promoter (RCAS-TV-A, Federspiel et al., 1994). Transgenic lines have been developed expressing the tv-a receptor from the mouse melanoblast promoter, dopachrome tautomerase.  In utero infection at E12.5 with an RCAS-Tyrosinase virus resulted in complementation of the albino (Tyrc) mutation in melanoblasts as evidenced by pigmented hairs after birth. A neural crest directed tv-a transgenic line was developed using the PAX3 promoter.  Neural tube explants isolated from the neural crest mutant Sox10lacZ/Sox10lacZ; PAX3-tv-a were complemented by RCAS-Sox10 virus in tissue explants.  These studies demonstrate the validity of using this approach as a rapid, functional test of genes for complementing genetic defects in a lineage directed manner.


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