STOCK Tg(Csf1r*-GAL4/VP16,UAS-ECFP)1Hume/J

Stock number: 026051
Product name: MacBlue
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

MacBlue transgenic mice may be useful as marker for the study of monocyte and dendritic cell trafficking and mononuclear phagocyte development in tissues.

Detailed description

MacBlue transgenic mice express enhanced cyan fluorescent protein (ECFP) from a truncated colony stimulating factor 1 receptor (Csf1r) promoter using the binary yeast Gal4-UAS system. The Gal4-UAS system utilizes a transgene that consists of both Gal4-expressing and Gal4-reporting modules to drive the expression of the ECFP reporter in Csf1r-expressing cells. In this strain, the Csf1r promoter lacks the 150 bp trophoblast and osteoclast-specific transcription start sites, directing reporter expression specifically in macrophages. Hemizygotes are viable and fertile. All ECFP⁺ cells in peripheral blood of MacBlue mice are CD11b-positive, and are negative for lymphocyte markers. 10% of granulocytes in peripheral blood and bone marrow express ECFP, as do Ly6C+ and Ly6C- monocytes. All ECFP^hi leukocytes express the monocyte/macrophage marker F4/80. 7% of bone marrow cells are ECFP⁺. ECFP was also detected in adult dendritic cells and blood monocytes in the gut, microglia and Langerhans cells, Peyer’s patches and isolated lymphoid follicles.

These mice may be bred to mice carrying tagged proteins under control of the Gal4-dependent promoter to direct expression of that protein in mononuclear phagocytes.

Development

MacBlue mice contain 2 cointegrated transgenes. One transgenic construct was designed with a colony stimulating factor 1 receptor (Csf1r) promoter, lacking the 150 bp trophoblast and osteoclast-specific transcription start sites. This mutated Csf1r drives expression of a Gal4 driver (containing a yeast gal4 transcriptional activator fused to herpes simplex virus protein (VP16)). A Gal4-responder plasmid was designed to contain an enhanced cyan fluorescent protein (ECFP) fused to yeast upstream activating sequence (UAS) Gal4-dependent promoter. The two constructs were coinjected into (C57BL6/JxCBA)F2 oocytes and founder mice were bred to CD1 outbred mice to establish a colony. Upon arrival at The Jackson Laboratory, transgenic mice were bred to C57BL/6J inbred mice (Stock No. 000664) to establish the colony.

Allele

Symbol: Tg(Csf1r*-GAL4/VP16,UAS-ECFP)1Hume
Name: transgene insertion 1, David A Hume
MGI accession ID: MGI:5587942
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Csf1r*-GAL4/VP16,UAS-ECFP)1Hume
Marker name: transgene insertion 1, David A Hume
Chromosome: UN
Strain of origin: (C57BL/6J x CBA)F2
Expressed genes: CFP,Cyan Fluorescent Protein,jellyfish; GAL4,transcriptional activator GAL4,yeast
Site of expression: GAL4 and ECFP are expressed in macrophages.
Molecular note: Two transgenes inserted at a single locus. One transgenic construct is designed with a colony stimulating factor 1 receptor (Csf1r) promoter, lacking the 158 bp (-454 to -298 relative to ATG) trophoblast transcription start sites and a TATA-box. This mutated Csf1r drives expression of a "Gal4 driver" (containing a yeast GAL4 transcriptional activator fused to herpes simplex virus protein (VP16)). A "Gal4-responder" plasmid is designed to contain an enhanced cyan fluorescent protein (ECFP) uder the control of a yeast upstream activating sequence (UAS) Gal4-dependent promoter. The founder line with approximately 5% ECFP+ leukocytes was selected for characterization.