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Cell type-specific expression of the human renin gene
A Fukamizu1, S Uehara, K Sugimura
1Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.
Summary
Transgenic mice carrying the human renin gene show kidney-specific expression, independent of gene copy number. This cell-type specific expression was confirmed in vitro, highlighting the regulatory elements of the renin gene.
Area of Science:
- Molecular Biology
- Genetics
- Physiology
Background:
- Previous production of transgenic mice with a human renin gene.
- Need for further characterization of tissue-specific gene expression.
Purpose of the Study:
- To characterize the tissue-specific expression of a human renin transgene in mice.
- To investigate the influence of transgene copy number on expression patterns.
- To confirm cell-type specific expression in vitro.
Main Methods:
- Northern blot analysis to detect human renin gene expression in transgenic mouse tissues (kidney, liver).
- Immunohistochemistry using a human renin-specific monoclonal antibody.
- In vitro transfection experiments with chloramphenicol acetyltransferase (CAT) reporter gene assays using kidney-derived (293) and liver-derived (HepG2) cell lines.
Main Results:
- Human renin gene expression was detected exclusively in the kidney, not the liver, of transgenic mice, irrespective of transgene copy number (10 or 50 copies).
- Immunohistochemistry confirmed human renin expression localized to epithelioid juxtaglomerular cells in the mouse kidney.
- In vitro transfection demonstrated activation of a renin gene promoter-driven CAT gene in kidney-derived 293 cells but not in liver-derived HepG2 cells.
Conclusions:
- The human renin gene exhibits dominant-renal and cell-type specific expression in transgenic mice.
- Transgene copy number does not appear to influence the tissue-specific expression pattern.
- In vitro studies support the presence of regulatory elements responsible for cell type-specific expression of the renin gene.