Exonuclease Epigenetic Sequencing

核酸外切酶表观遗传测序

• 批准号: 10009454
• 负责人: Eric Ervin
• 金额: $ 89.7万
• 依托单位: ELECTRONIC BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10009454
• 关键词: Affinity; Biological Sciences; Biology; Chemicals; Cleaved cell; Collaborations; Communicable Diseases; Communities; Computer software; Contracts; Coronary heart disease; Custom; DNA; DNA sequencing; Data; Detection; Development; Diabetes Mellitus; Diagnostic; Disease; Electrodes; Electronics; Ensure; Epigenetic Process; Escherichia coli; Exonuclease; Explosion; Gene Expression; Genetic Markers; Genome; Genomic DNA; Genomics; Goals; Gold; Growth; Hemolysin; Heritability; High-Throughput Nucleotide Sequencing; Individual; Infection; Information Systems; Laboratories; Lipid A; Lipid Bilayers; Malignant Neoplasms; Methodology; Methods; Modification; Mutagenesis; Mutate; New England; Noise; Nucleic Acids; Nucleotides; Phase; Production; Publishing; Resolution; Role; Sampling; Single-Stranded DNA; Stretching; Sugar Phosphates; Techniques; Technology; Therapeutic; Time; Validation; Vertebral column; Viscosity; base; cancer therapy; cell free DNA; clinical diagnostics; data acquisition; design; diagnostic biomarker; empowered; epigenetic marker; epigenome; epigenomics; infancy; instrument; instrumentation; multidisciplinary; mutant; nanopore; nervous system disorder; new technology; next generation sequencing; novel; pathogen; prognostic; programs; prototype; response; sensor; sequencing platform; solid state; success; synthetic construct; temporal measurement; tool

Project Summary There is a current need within the field of next generation sequencing (NGS) for new, enabling instrumentation, capable of high accuracy, direct, native DNA sequencing, including the identification of canonical and modified bases and the correct characterization of homopolymer stretches and repeating sequences. During this program, Electronic BioSciences (EBS) and a team of field experts aim to solve the technical challenges associated with the development of a completely new and novel nanopore-based sequencing platform, including the associated methodology for sequencing DNA at the single nucleotide level, with the capability of directly and correctly identifying chemically modified nucleotides. During this project, efforts will specifically focus on the high accuracy detection/identification of 5-methylcytosine (5mC) and N6-methyladenine (m6A) sequencing, before pursuing other modifications. At present, the scientific community’s understanding of the “epigenome,” i.e. the chemical modifications which regulate the function of DNA, is still in its infancy. While there are many known chemical modifications to either the base or sugar-phosphate backbone of nucleic acids, due to the lack of analytical characterization methods available, the exact roles of these modifications remain to be assessed. New technologies capable of elucidating the roles of these modifications have the potential to revolutionize the use of the epigenome. Furthermore, with regards to homopolymer and repeat sequences of >6 nucleotides which are commonly found in genomes, clinicians have identified that many of these sequences are expanded, contracted, or mutated in cancers, neurological disorders, and heritable diseases, and therefore, sequencing for changes in these regions has promising potential for utilization as routine genetic markers for diagnostics and prognostics purposes. At the conclusion of this Fast-Track project, a multiplexed sequencing instrument that is ready for immediate, expanded, initial user base use in laboratory settings will be developed and built, and complete concept feasibility will have been demonstrated through both synthetic and genomic DNA sequencing, including 5mC and m6A characterization.

项目摘要 在下一代测序（NGS）的领域中，当前需要新的，启用仪器， 能够高精度，直接的天然DNA测序，包括识别规范和修改 均聚物的碱和正确表征伸展和重复序列。在此程序中， 电子生物科学（EBS）和一个田野专家团队旨在解决与之相关的技术挑战 一个全新新颖的基于纳米孔的测序平台的开发，包括相关的 在单个核苷酸水平上测序DNA的方法，直接，正确的能力 鉴定化学修饰的核苷酸。在此项目中，努力将特别关注高精度 在追求之前，检测/鉴定5-甲基环霉素（5MC）和N6-甲基丹宁（M6A）测序 其他修改。目前，科学界对“表观基因组”的理解，即化学 调节DNA功能的修饰仍处于起步阶段。虽然有许多已知的化学物质 由于缺乏分析，对核酸的碱基或糖磷酸主链修饰 可用的表征方法，这些修饰的确切作用仍有待评估。新的 能够阐明这些修饰作用的技术有可能革新使用 表观基因组。此外，关于均聚物和> 6个核苷酸的重复序列 临床医生通常在基因组中发现，其中许多序列都扩展，收缩， 或在癌症，神经系统疾病和可遗传疾病中发生突变，因此，对变化进行了测序 这些区域已承诺将利用作为诊断和预后的常规遗传标记的潜力 目的。在这个快速轨道项目的结论中 将开发和构建并完整的实验室设置中的立即，扩展的初始用户群使用 概念可行性将通过合成和基因组DNA测序证明 5MC和M6A表征。