2013 Fiscal Year Final Research Report

Construction of cell-culture based assay system to understand CGG repeat instability in Fragile X syndrome

Research Project

Project/Area Number	24770005
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Genetics/Genome dynamics
Research Institution	Tottori University
Principal Investigator	NAKAYAMA Yuji 鳥取大学, 生命機能研究支援センター, 助教 (40432603)
Project Period (FY)	2012-04-01 – 2014-03-31
Keywords	脆弱X症候群 / トリプレットリピート / 染色体工学 / 細胞融合
Research Abstract	The human Fragile X mental retardation 1 (FMR1) gene, which contains CGG-trinucleotide repeat in its 5'UTR region, is responsible gene for Fragile X syndrome (FXS). The number of CGG-repeat is about 50 to 200 in carriers (so-called 'premutation allele') and more in patients. CGG-repeat becomes unstable and in most cases expands upon maternal inheritance of premutation allele to children. However, the mechanism underling CGG-repeat instability is unknown to date. In the present study, toward elucidation the molecular and cellular mechanism in CGG-repeat instability, we tried to establish a cell culture-based system that enables to reproduce CGG-repeat instability in vitro. We applied chromosomal engineering so that we could clone and handle CGG-repeat tract as a chromosomal domain stably. We successfully cloned natural and artificial chromosomes that contain varying size of CGG-repeats in rodent cell lines.