Genome-wide methylation pattern search in Prader-Willi Syndrome (PWS) patients

• Main Authors: Ronald-Garingalao Garvilles, 蓋羅納
• Other Authors: Chung-Yung Chen
• Format: Others
• Language: en_US
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/59827189279715437068

碩士 === 中原大學 === 生物科技研究所 === 99 === Prader-Willi Syndrome (PWS) is a neurodevelopmental disorder caused by lack of functional paternal copy of 15q11-q13. Epigenetic aberrations due to genomic imprinting defect leads to the absence expression of paternally-inherited genes. This study aims to provide an overview of the methylation status of the whole-genome among PWS patients. A novel method PCR Selective Suppression Hybridization (PSSH) was employed to screen the putative genes that are differentially methylated among PWS patients but not in normal individuals. This method utilizes hybridization of methylated-non-biotin labeled DNA and unmethylated-biotin-labeled DNA from the PWS patient and mother, respectively, after subsequent bisulfite treatment, USER enzyme cleavage, labeling with biotin dCTP and separation by streptavidin. After successful screening of two families, several genes and gene clusters were identified that could possibly link to PWS after comprehensive analyses using Database for Annotation, Visualization and Integrated Discovery (DAVID) and Pathway Interaction Database (PID): CREBBP, RRM1, HPP1 and XRCC6. Based on the previous published results, these candidate genes were found to have cross-talk mechanisms that link between DNA methylation and histone covalent modifications. From the confirmed PWS-related genes based on OMIM (NCBI), co-regulation of androgen receptor activity and E2F transcription factor network pathways could be candidate related pathways on the screened putative genes. In order to establish these hypothetical results, more PWS samples are needed for the analysis; and will be confirmed using real-time PCR (qPCR), methylation specific PCR (MSP) and bisulfite sequencing PCR (BSP).