A novel homologous recombination system to study 92 kDa type IV collagenase transcription demonstrates that the NFκB motif drives the transition from a repressed to an activated state of gene expression

Abstract
SPECIFIC AIMSThe 92 kDa type IV collagenase (MMP-9) plays a major role in physiological and pathological events such as trophoblast implantation in pregnancy, bone development, and tumor progression. Regulation of MMP-9 protein levels has been largely ascribed to transcriptional activation of the gene. Considering its role in physiology and pathology, there is a need for an increased understanding of how this gene is transcriptionally controlled. In the present study, we report on the development and characterization of a novel system (which overcomes several limitations associated with studies using extrachromosomal plasmids) in which a single copy of a wild-type or mutated MMP-9 promoter fragment(s) fused to a luciferase reporter was inserted at a Flp recombinase target site (FRT) in the HT1080 cell genome.PRINCIPAL FINDINGSTo direct MMP-9 promoter-reporter constructs to a single genomic locus, we first generated constructs bearing a MMP-9 regulatory sequence-driven luciferase cDNA and a Flp FRT site. T...
Funding Information
  • Dutch Dairymen Board (R01 CA89002, R01 DE 10845, P50‐CA097007)

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