title: Complementation of melanocyte development in SOX10 mutant neural crest using lineage-directed gene transfer.
abstract: An in vitro gene complementation approach has been developed to dissect gene function and regulation in neural crest (NC) development and disease. The approach uses the avian RCAS virus to express genes in NC cells derived from transgenic mice expressing the RCAS receptor TVA, under the control of defined promoter elements. Constructs for creating TVA transgenic mice were developed using site-specific recombination GATEWAY (GW), compatible vectors that can also be used to facilitate analysis of genomic fragments for transcriptional regulatory elements. By using these GW vectors to facilitate cloning, transgenic mouse lines were generated that express TVA in SOX10-expressing NC stem cells under the control of the Pax3 promoter. The Pax3-tv-a transgene was bred onto a Sox10-deficient background, and the feasibility of complementing genetic NC defects was demonstrated by infecting the Pax3-tv-a cells with an RCAS-Sox10 expression virus, thereby rescuing melanocyte development of Sox10-deficient NC cells. This system will be useful for assessing genetic hierarchies in NC development. Developmental Dynamics 229:54-62, 2004.
authors.affiliation: Mouse Embryology Section, Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Ling
authors.initials: L
authors.lastname: Hou
authors.forename: Stacie K
authors.initials: SK
authors.lastname: Loftus
authors.forename: Arturo
authors.initials: A
authors.lastname: Incao
authors.forename: Amy
authors.initials: A
authors.lastname: Chen
authors.forename: William J
authors.initials: WJ
authors.lastname: Pavan
mesh_terms: D000818:Animals; D030161:Avian Proteins; D001483:Base Sequence; D002454:Cell Differentiation; D004247:DNA / Q000235:genetics; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics*; D018014:Gene Transfer Techniques; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D006609:High Mobility Group Proteins / Q000172:deficiency / Q000235:genetics*; D066298:In Vitro Techniques; D008544:Melanocytes / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009154:Mutation; D009432:Neural Crest / Q000166:cytology* / Q000378:metabolism*; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D011401:Promoter Regions, Genetic; D011991:Receptors, Virus / Q000235:genetics; D055757:SOXE Transcription Factors; D014157:Transcription Factors
chemical_list: D030161:Avian Proteins; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D000072041:PAX3 Transcription Factor; D051761:Paired Box Transcription Factors; D011991:Receptors, Virus; D055757:SOXE Transcription Factors; C527924:Sox10 protein, mouse; D014157:Transcription Factors; C088745:Tva receptor; C068686:Pax3 protein, mouse; D004247:DNA