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 当前在下一代测序（NGS）领域内需要新的、使能的仪器， 能够高准确性，直接，天然DNA测序，包括鉴定典型和修饰的 碱基和均聚物延伸和重复序列的正确表征。在这个节目中， Electronic BioSciences（EBS）和一个现场专家团队旨在解决与 开发全新的基于纳米孔的测序平台，包括相关的 在单核苷酸水平上对DNA进行测序的方法学，具有直接和正确地 识别化学修饰的核苷酸。在这个项目中，工作将特别集中在高精度 5-甲基胞嘧啶（5 mC）和N6-甲基腺嘌呤（m6 A）测序的检测/鉴定， 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 由于缺乏对核酸的碱基或糖-磷酸骨架的分析性修饰， 尽管现有的表征方法不多，但这些修饰的确切作用仍有待评估。新 能够阐明这些修饰的作用的技术有可能彻底改变 表观基因组此外，关于多6个核苷酸的均聚物和重复序列， 通常在基因组中发现，临床医生已经确定，这些序列中的许多是扩展的，收缩的， 或在癌症、神经系统疾病和遗传性疾病中发生突变，因此， 这些区域具有作为诊断和鉴别的常规遗传标记的应用潜力 目的在这个快速通道项目结束时，一个多路测序仪器，准备用于 将开发和建立并完成在实验室环境中使用的即时、扩大的初始用户基础 概念的可行性将通过合成和基因组DNA测序得到证明，包括 5 mC和m6 A表征。