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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.

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  • Authors+Show Affiliations

    ,

    Institute for Molecular Bioscience, Bldg. 80, University of Queensland, Brisbane, Queensland 4072, Australia. d.ovchinnikov@imb.uq.edu.au

    , , , ,

    Source

    Journal of leukocyte biology 83:2 2008 Feb pg 430-3

    MeSH

    Animals
    Cell Line
    Cell Lineage
    Crosses, Genetic
    DNA-Binding Proteins
    Dendritic Cells
    Gene Expression Regulation, Developmental
    Genes, Reporter
    Genes, Synthetic
    Genes, fms
    Green Fluorescent Proteins
    Macrophages
    Mice
    Mice, Inbred C57BL
    Mice, Inbred CBA
    Mice, Transgenic
    Microglia
    Monocytes
    Organ Specificity
    Promoter Regions, Genetic
    Saccharomyces cerevisiae Proteins
    Transcription Factors
    Transcriptional Activation
    Transgenes
    Yolk Sac

    Pub Type(s)

    Journal Article
    Research Support, Non-U.S. Gov't

    Language

    eng

    PubMed ID

    17971498

    Citation

    Ovchinnikov, Dmitry A., et al. "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." Journal of Leukocyte Biology, vol. 83, no. 2, 2008, pp. 430-3.
    Ovchinnikov DA, van Zuylen WJ, DeBats CE, et al. 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. J Leukoc Biol. 2008;83(2):430-3.
    Ovchinnikov, D. A., van Zuylen, W. J., DeBats, C. E., Alexander, K. A., Kellie, S., & Hume, D. A. (2008). 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. Journal of Leukocyte Biology, 83(2), pp. 430-3.
    Ovchinnikov DA, et al. 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. J Leukoc Biol. 2008;83(2):430-3. PubMed PMID: 17971498.
    * Article titles in AMA citation format should be in sentence-case
    TY - JOUR T1 - 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. AU - Ovchinnikov,Dmitry A, AU - van Zuylen,Wendy J M, AU - DeBats,Claire E E, AU - Alexander,Kylie A, AU - Kellie,Stuart, AU - Hume,David A, Y1 - 2007/10/30/ PY - 2007/11/1/pubmed PY - 2008/4/10/medline PY - 2007/11/1/entrez SP - 430 EP - 3 JF - Journal of leukocyte biology JO - J. Leukoc. Biol. VL - 83 IS - 2 N2 - 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. SN - 0741-5400 UR - https://www.unboundmedicine.com/medline/citation/17971498/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_ L2 - https://doi.org/10.1189/jlb.0807585 DB - PRIME DP - Unbound Medicine ER -