title: Development and application of a new Silent reporter system to quantitate the activity of enhancer elements in the type II Collagen Gene.
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.
authors.affiliation: 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.
authors.forename: Kazuo
authors.initials: K
authors.lastname: Ito
authors.affiliation: Tissue Regeneration, Department of Bio-Matrix, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8549, Japan. Electronic address: t.shinomura.trg@tmd.ac.jp.
authors.forename: Tamayuki
authors.initials: T
authors.lastname: Shinomura
mesh_terms: D053674:Aggrecans / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D002356:Cartilage / Q000378:metabolism; D045744:Cell Line, Tumor; D019902:Chondrocytes / Q000378:metabolism; D024043:Collagen Type II / Q000096:biosynthesis / Q000235:genetics*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005786:Gene Expression Regulation / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: D053674:Aggrecans; D024043:Collagen Type II
keywords: Aggrecan; Cartilage; Chondrocytes; Enhancer; Promoter; Type II collagen