title: Expression of Gal4-dependent transgenes in cells of the mononuclear phagocyte system labeled with enhanced cyan fluorescent protein using Csf1r-Gal4VP16/UAS-ECFP double-transgenic mice.
abstract: We generated double-transgenic mice carrying cointegrated tissue-specific Gal4 and Gal4 reporter transgenes to direct transgene overexpression in the mononuclear phagocyte system (MPS). A modified promoter of the Csf1r (c-fms) gene, containing a deletion of the trophoblast-specific promoter, was used to drive the expression of Gal4VP16 transcriptional activator specifically in macrophages. This module was cointegrated with a fluorescent reporter, enhanced cyan fluorescent protein (ECFP), driven by a Gal4-dependent promoter. ECFP fluorescence was first detected in forming blood islands of the yolk sac at 8 dpc, then in macrophages in the yolk sac and the embryo proper. In adult mice ECFP was detected primarily in monocytes, tissue macrophages, microglia, and dendritic cells, including Langerhans cells of the skin. Crossing of these mice to transgenics containing tagged protein under control of a Gal4-dependent promoter directed expression of that protein in mononuclear phagocytes of double-transgenic animals. The new mouse line provides a useful tool for overexpression of transgenes in cells of the myeloid lineage, while simultaneously labeling them by ECFP expression.
authors.affiliation: Institute for Molecular Bioscience, Bldg. 80, University of Queensland, Brisbane, Queensland 4072, Australia. d.ovchinnikov@imb.uq.edu.au
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Ovchinnikov
authors.forename: Wendy J M
authors.initials: WJ
authors.lastname: van Zuylen
authors.forename: Claire E E
authors.initials: CE
authors.lastname: DeBats
authors.forename: Kylie A
authors.initials: KA
authors.lastname: Alexander
authors.forename: Stuart
authors.initials: S
authors.lastname: Kellie
authors.forename: David A
authors.initials: DA
authors.lastname: Hume
mesh_terms: D000818:Animals; D002460:Cell Line / Q000378:metabolism; D019070:Cell Lineage; D003433:Crosses, Genetic; D004268:DNA-Binding Proteins; D003713:Dendritic Cells / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D005813:Genes, Synthetic / Q000235:genetics*; D016236:Genes, fms; D049452:Green Fluorescent Proteins / Q000235:genetics*; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic / Q000196:embryology / Q000235:genetics*; D017628:Microglia / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism*; D009928:Organ Specificity; D011401:Promoter Regions, Genetic / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics*; D015533:Transcriptional Activation; D019076:Transgenes / Q000235:genetics; D015017:Yolk Sac / Q000166:cytology
chemical_list: D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C527701:enhanced cyan fluorescent protein; D049452:Green Fluorescent Proteins