High-throughput sequencing of chromatin regulatory elements

ENCODE联盟项目旨在系统地识别和分类人类基因组中的染色质调控元件。这一目标正在通过应用染色质方法和超高通量测序来实现，以获得反映组蛋白修饰和染色质蛋白在给定细胞类型中的分布的全基因组图谱。通过分析多种细胞类型，该项目将建立一个越来越全面的染色质调控元件目录，并深入了解它们的细胞类型特异性。尽管染色质中有几十种不同的修饰和许多其他结构成分，但只有一小部分可以用现有的抗体和试剂来测量。这一补充要求的基本原理是，当这些修饰和蛋白质在基因组中发生时，表征这些修饰和蛋白质的能力将增强我们对ENCODE所阐明的功能元件的结构和调控的理解，并揭示未知意义的其他元件。申请的资金将用于筛选数十个额外的染色质表位，目的是确定适合全基因组定位的试剂和方法。筛选将通过常规方法和高通量技术相结合进行。通过这一过程鉴定的试剂将被应用于现有的ENCODE管道中，以表征新表位的全基因组分布及其与已知或新的调控元件的关系。

This ENCODE consortium project aims to identify and classify chromatin regulatory elements in the human genome in systematic fashion. This goal is being pursued through the application of chromatin methods and ultra high-throughput sequencing to acquire genomewide maps that reflect the distributions of histone modifications and chromatin proteins in a given cell type. By assaying multiple cell types, the project will build an increasingly comprehensive catalog of chromatin regulatory elements and gain insight into their cell typespecificities. Although there are dozens of different modifications and many other structural components in chromatin, only a fraction can be measured with existing antibodies and reagents. The rationale of this supplement request is that an ability to characterize these additional modifications and proteins as they occur across the genome would augment our understanding of the structure and regulation of functional elements elucidated by ENCODE, and reveal additional elements of yet unknown significance. Requested funds would be use to screen dozens of additional chromatin epitopes with the goal of identifying suitable reagents and methods for genomewide mapping. The screens will be pursued through a combination of conventional methods and high-throughput techniques. Reagents identified through this process will then be applied within the existing ENCODE pipeline to characterize genomewide distributions of the novel epitopes and their relationships to known or novel regulatory elements. Chromatin regulation is of wide-ranging importance in human development and disease. The proposed systematic identification and characterization of chromatin regulatory elements in the human genome will offer valuable insight into the structure and function of chromatin, and provide a resource for investigators in chromatin, genomics, cancer and many other fields of research. All data collected in the context of the proposed project will be made available pre-publication to the greater scientific community as soon as reliability has been confirmed. 2.