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The myeloperoxidase gene proximal enhancer directs hematopoietic-specific expression in transgenic mice
1Department of Molecular Biology, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, NJ, USA.
Abstract:
The myeloperoxidase (MPO) gene is expressed specifically in immature myeloid cells. The MPO gene includes a promoter proximal enhancer which is coincident with DNaseI hypersensitive chromatin sites and is specifically active in myeloid cell lines. We developed transgenic murine lines in which 1.3 kb of murine MPO proximal 5' flanking region DNA was linked to a TATAA homology and RNA initiation site derived from the HSV-TK promoter and to a luciferase reporter (MPOTKLUC). In each of six founder lines, high-level luciferase activity was evident in marrow, thymus and spleen. Modest- to high-level luciferase expression was also evident in brain and in the heart in several of the lines, and luciferase activity was at or near background levels in lung, liver, kidney, stomach, colon, bladder, skeletal muscle, skin and small intestine in all of the MPOTKLUC transgenic mice. Within marrow cells, luciferase activity was evident in myeloid (GR-1+), B lymphoid (B220+) and T-lymphoid (CD4+) cells. Additional regulatory regions, thus, may be required to further restrict MPO gene expression to immature myeloid cells.
Insights
The myeloperoxidase (MPO) gene
Area of Science:
- Molecular Biology
- Genetics
- Immunology
Background:
- The myeloperoxidase (MPO) gene is specifically expressed in immature myeloid cells.
- A promoter proximal enhancer in the MPO gene is associated with DNaseI hypersensitive chromatin sites and is active in myeloid cell lines.
Purpose of the Study:
- To investigate the regulatory elements controlling MPO gene expression.
- To develop transgenic mouse models for studying MPO gene regulation.
Main Methods:
- Developed transgenic murine lines (MPOTKLUC) by linking 1.3 kb of murine MPO 5' flanking DNA to an HSV-TK promoter and luciferase reporter.
- Analyzed luciferase activity in various tissues and cell types of the transgenic mice.
Main Results:
- High-level luciferase expression was observed in marrow, thymus, and spleen of all founder lines.
- Modest- to high-level expression was also detected in brain and heart in several lines.
- Luciferase activity was low in other tissues, and within marrow cells, expression was found in myeloid, B lymphoid, and T-lymphoid cells.
Conclusions:
- The 1.3 kb MPO 5' flanking region drives gene expression in multiple tissues and cell types.
- Additional regulatory elements are likely necessary to restrict MPO expression specifically to immature myeloid cells.
