IMR Press / FBL / Volume 7 / Issue 1 / DOI: 10.2741/A751

Frontiers in Bioscience-Landmark (FBL) is published by IMR Press from Volume 26 Issue 5 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on as a courtesy and upon agreement with Frontiers in Bioscience.

Promoter competition assay for analyzing gene regulation in joint tissue engineering
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1 Biomedical Engineering Program, Departments of Anatomy and Cell Biology, and of Mechanical Engineering, Indiana University - Purdue University Indianapolis, Indianapolis, Indiana 46202
2 Department of Biology, Indiana University, Bloomington, Indiana 47405
Front. Biosci. (Landmark Ed) 2002, 7(1), 169–174;
Published: 1 August 2002

We describe a new biochemical technique, "promoter competition assay," for examining the role of cis-acting DNA elements in tissue cultures. Recent advances in tissue engineering permit the culture of a variety of cells. Many tissues are engineered, however, without an appropriate understanding of molecular machinery that regulates gene expression and cellular growth. For elucidating the role of cis-acting regulatory elements in cellular differentiation and growth, we developed the promoter competition assay. This assay uses a transient transfer into cells of double-stranded DNA fragments consisting of cis-acting regulatory elements. The transferred DNA fragments act as a competitor and titrate the function of their genomic counterparts. Using synovial cells derived from a rheumatoid arthritis patient, we examined a role of NF-kB binding sites in the regulation of the expression of matrix metalloproteinase (MMP) genes. The results support a stimulatory role of NF-kB in transcriptional regulation of MMP-1 and MMP-13.

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