Abstract
Although the promoter/enhancer of the IL-2 gene mediates inducible reporter gene expression in vitro, it cannot drive consistent expression in transgenic mice. The location and existence of any regulatory elements that could open the IL-2 locus in vivo have remained unknown, preventing analysis of IL-2 regulation in developmental contexts. In this study, we report the identification of such a regulatory region, marked by novel DNase-hypersensitive sites upstream of the murine IL-2 promoter in unstimulated and stimulated T cells. Inclusion of most of these sites in an 8.4-kb IL-2 promoter green fluorescent protein transgene gives locus control region-like activity. Expression is efficient, tissue specific, and position independent. This transgene is expressed not only in peripheral T cells, but also in immature thymocytes and thymocytes undergoing positive selection, in agreement with endogenous IL-2 expression. In contrast, a 2-kb promoter green fluorescent protein transgene, lacking the new hypersensitive sites, is expressed in only a few founder lines, and expression is dysregulated in CD8(+) cells. Thus, the 6.4 kb of additional upstream IL-2 sequence contains regulatory elements that provide integration site independence and differential regulation of transgene expression in CD8 vs CD4 cells.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3' Untranslated Regions / immunology
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5' Untranslated Regions / immunology
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Animals
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Base Composition / immunology
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CD8-Positive T-Lymphocytes / immunology
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CD8-Positive T-Lymphocytes / metabolism
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Cell Lineage / genetics
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Cell Lineage / immunology
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Cell Separation
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Cells, Cultured
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Deoxyribonuclease I / genetics
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Gene Dosage
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Gene Expression Regulation / immunology*
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Gene Expression Regulation, Developmental / immunology
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Genetic Markers / immunology
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Genetic Vectors / chemical synthesis
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Genetic Vectors / immunology
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Green Fluorescent Proteins
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Immunologic Memory / genetics
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Immunophenotyping
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Interleukin-2 / biosynthesis*
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Interleukin-2 / genetics*
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Luminescent Proteins / biosynthesis
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Luminescent Proteins / genetics
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Lymphocyte Activation / genetics
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Male
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Mice
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Mice, Inbred C57BL
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Mice, Inbred DBA
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Mice, SCID
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Mice, Transgenic
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Molecular Sequence Data
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Receptors, Antigen, T-Cell, alpha-beta / genetics
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Regulatory Sequences, Nucleic Acid / immunology*
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Response Elements / immunology
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T-Lymphocyte Subsets / immunology
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T-Lymphocyte Subsets / metabolism
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Transgenes / immunology*
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Tumor Cells, Cultured
Substances
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3' Untranslated Regions
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5' Untranslated Regions
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Genetic Markers
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Interleukin-2
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Luminescent Proteins
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Receptors, Antigen, T-Cell, alpha-beta
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Green Fluorescent Proteins
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Deoxyribonuclease I