Comprehensive Single Molecule Enhanced Detection of Modified Cytosines in Mammalian Genomes

哺乳动物基因组中修饰胞嘧啶的综合单分子增强检测

• 批准号: 9316069
• 负责人: TIMOTHY H BESTOR
• 金额: $ 32万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-19 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9316069
• 关键词: Binding; Binding Sites; Biological Process; Catalytic Domain; Charge; Chemicals; Chemistry; CpG dinucleotide; Cytosine; DNA; DNA Methylation; DNA Modification Process; DNA sequencing; Detection; Dioxygenases; Disease; Enzymatic Biochemistry; Enzymes; Epigenetic Process; Gene Expression Profile; Genes; Genome; Genomics; Human; Human Genome; Label; Lead; Malignant Neoplasms; Maps; Mediating; Methionine; Methods; Methylation; Methyltransferase; Modeling; Modification; Noise; Nucleotides; Oxides; Pattern; Positioning Attribute; Promoter Regions; Protocols documentation; Resolution; S-Adenosylmethionine; Side; Signal Transduction; Site; Site-Directed Mutagenesis; Technology; Testing; Time; Ursidae Family; analog; base; bisulfite sequencing; cancer genome; cofactor; genome wide methylation; glucosyltransferase B; human disease; improved; mammalian genome; methyl group; methylation pattern; mutant; nanopore; novel; novel strategies; sequencing platform; single molecule; solid state; sugar; whole genome

Project abstract Patterns of modified cytosines (5-methylcytosine and 5-hydroxymethylcytosine) are required for the normal function of the genome, and perturbations of patterns of modified cytosine can be lethal and lead to gross changes in patterns of gene expression. Methylation patterns are grossly abnormal in many forms of human cancer. Existing methods of genome-wide methylation profiling require large amounts of DNA, overestimate methylation levels, and cannot cover the entire genome. We are developing a method wherein enzymes are used to modify sites of methylation and hydroxymethylation with chemical tags that render these sites easily detectible by nanopore sequencing by synthesis and other platforms for single molecule DNA sequencing. The result will be a comprehensive method for the detection of modified bases in human genomes with much greater sensitivity, accuracy, economy, and throughput than existing methods.

项目摘要 修饰胞嘧啶（5-甲基胞嘧啶和5-羟甲基胞嘧啶）的模式 是基因组正常功能所必需的， 修饰的胞嘧啶可能是致命的，并导致基因模式的巨大变化 表情甲基化模式在许多形式的人类中是严重异常的， 癌现有的全基因组甲基化谱分析方法需要大量的 大量的DNA，高估了甲基化水平，不能覆盖整个 基因组我们正在开发一种方法， 甲基化和羟甲基化的位点与化学标签，使这些 通过合成和其他平台的纳米孔测序容易检测的位点 用于单分子DNA测序。其结果将是一个全面的方法 用于检测人类基因组中的修饰碱基， 灵敏度、准确度、经济性和生产量。