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Development and application of a new Silent reporter system to quantitate the activity of enhancer elements in the type II Collagen Gene.

Authors
  • Ito, Kazuo1
  • Shinomura, Tamayuki2
  • 1 Tissue Regeneration, Department of Bio-Matrix, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8549, Japan; Department of Orthopaedic Surgery, Sendai Teishin Hospital, Sendai, Miyagi 980-8798, Japan. , (Japan)
  • 2 Tissue Regeneration, Department of Bio-Matrix, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8549, Japan. Electronic address: [email protected]. , (Japan)
Type
Published Article
Journal
Gene
Publication Date
Jul 01, 2016
Volume
585
Issue
1
Pages
13–21
Identifiers
DOI: 10.1016/j.gene.2016.03.019
PMID: 26992640
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Type II collagen is a major component of cartilage, which provide structural stiffness to the tissue. As a sufficient amount of type II collagen is critical for maintaining the biomechanical properties of cartilage, its expression is tightly regulated in chondrocytes. Therefore, it is essential to elucidate in detail the transcriptional mechanism that controls expression of type II collagen, in particular by two enhancer elements we recently discovered. To systematically analyze and compare enhancer activities, we developed a novel reporter assay system that exploits site-specific integration of promoter and enhancer elements to activate a transcriptionally silent reporter gene. Using this system, we found that the enhancer elements have distinct characteristics, with one exhibiting additive effects and the other exhibiting synergistic effects when repeated in tandem. Copyright © 2016 Elsevier B.V. All rights reserved.

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