17 表观遗传学_染色质结构的分子水平研究201410.ppt

* 每次拉动1bp * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * 染色质结构变化是表观遗传调控的核心 Epigenetics chromatin Transcription Histone variants Chromatin remodeling Non-coding RNA DNA methylation Histone modifications Chromatin transactors Roeder, 2010 干细胞编程与重编程中的表观遗传调控网络 染色质成分及反式作用因子 组蛋白变体：H2A.Z，H3.3，macroH2A 组蛋白修饰：Ac，Me，Ub 染色质多梳蛋白复合物 (PRC1 and PRC2)，MBT，HP1， Lamin A/C 染色质调节因子 组蛋白分子伴侣: NAP1, FACT, NASP 染色质重塑因子: SWI/SNF, RSF, ACF DNA修饰: 5mC or 5hmC reprogramming differentiation ES Cell Section 4 Approaches DNA所处的环境：核小体 如何检测open chromatin ？ Isolation of nuclei (108 cells) DNaseI treatment Embedded DNA into agarose plug Determined distribution of DNA using PFGE Choose a few DNaseI concentrations according to distribution In vivo Enriched for fragments from HS on a streptavidin column Identify Open chromatin：DNaseI hypersensitivity assay In vitro Ligated biotinylated linkers to DNA fragments sonicated to shear DNA Crawford G et al. Nature Methods, 2006 Crawford G et al. Cold Spring Harb Protoc 2010 Added second linker, amplified, labeled and hybridized to tiling microarray 研究方法 研究方法：Chromatin conformatio capture Hagege et al. Nature protocols, 2008 研究方法：ChIP (Chromatin Immunoprecipitation) for protein binding Section 5 Applications High-resolution mapping and characterization of open chromatin across the genome. Cell, 2008. 132(2): 311-22. Open chromatin in pluripotency and reprogramming. Nat Rev Mol Cell Biol 12, 36-47 (2011). Chd1 regulates open chromatin and pluripotency of embryonic stem cells. Nature 460, 863-868 (2009). A map of open chromatin in human pancreatic islets. Nat Genet 42,3,255-259(2010). Epigenetic instability of cytokine and transcription factor gene loci underlies plasticity of the T helper 17 cell lineage. Immunity 32, 616–627 (2010). 基因组注释 重编程 胚胎干细胞 胰岛细胞 Th17细胞发育 * Application in monogenic disease Facioscapulohumeral muscular dystrophy (FSHD) One of the strangest known autosomal dominant diseases.