Nanoscale Tools for Inosine Sequencing

用于肌苷测序的纳米级工具

• 批准号: 10437956
• 负责人: Eric Ervin
• 金额: $ 100万
• 依托单位: ELECTRONIC BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10437956
• 关键词: Achievement; Adenosine; Age-Years; Algorithms; Alzheimer' s Disease; Benchmarking; Biological Sciences; Cause of Death; Cell Culture Techniques; Cells; Code; Communities; Computer software; Dementia; Detection; Development; Diagnostic; Disease; Disease Progression; Enzymes; Exonuclease; Foundations; Goals; Guanosine; Hemolysin; Human; Individual; Inosine; Libraries; Malignant Neoplasms; Messenger RNA; Methods; MicroRNAs; Modeling; Modification; Molecular; Monitor; Nucleotides; Persons; Phase; Preparation; Procedures; Process; Property; RNA; RNA Editing; RNA Splicing; RNA-targeting therapy; Reader; Reading; Research; Research Personnel; Resolution; Risk; Role; Sampling; Site; Small RNA; Societies; System; Techniques; Technology; Temperature; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Time; Transfer RNA; United States; Virus Diseases; Viscosity; Work; age related; base; base editing; cancer type; data acquisition; epitranscriptomics; high reward; high risk; improved; instrument; mutant; nanofabrication; nanopore; nanoscale; nerve stem cell; nervous system disorder; novel; prognostic; programs; prototype; sensor; sequencing platform; single molecule; success; technology development; therapy outcome; tool; transcriptome sequencing

Project Summary During this program, Electronic BioSciences (EBS) will fully develop and demonstrate a completely new RNA sequencing technology capable of directly sequencing the most common product of RNA editing: inosine (I). Cells diversify the coding potential of their mRNA and regulatory small RNAs by enzymatic conversion of adenosine (A) to I, i.e., A-to-I editing, in which the edited base codes differently in mRNA, guides alternative mRNA splicing, and renders miRNA and tRNA functional in cells. The current inability to directly sequence sites of A-to-I RNA editing with quantitative readout (i.e., profile or determine their ratio within given sequences) seriously limits the ability of researchers to fully understand this epitranscriptomic process in disease. Thus, EBS aims to fill this technology gap by developing a single-molecule sequencing technique capable of directly identifying the four canonical nucleotides along with I, all with high resolution and high accuracy. In turn, such a technology development will improve the understanding of RNA function, including the role and significance of I, to enable better diagnostics, prognostics, and therapeutics. At the end of this Phase II effort, EBS will have developed a beta-prototype I sequencing system and demonstrated its complete functionality and analytical capabilities for a variety of RNA types as well as benchmarked the sequencing results against current state-of-the-art approaches.

项目概要 在此计划期间，Electronic BioSciences (EBS) 将全面开发并展示一种全新的 RNA 测序技术能够直接对最常见的RNA编辑产物：肌苷（I）进行测序。 细胞通过酶促转化使其 mRNA 和调节性小 RNA 的编码潜力多样化 腺苷 (A) 到 I，即 A 到 I 编辑，其中编辑的碱基在 mRNA 中编码不同，指导替代方案 mRNA 剪接，并使 miRNA 和 tRNA 在细胞中发挥功能。目前无法直接测序 具有定量读出功能的 A-to-I RNA 编辑位点（即，分析或确定它们在给定范围内的比率） 序列）严重限制了研究人员充分理解这一表观转录组过程的能力 疾病。因此，EBS旨在通过开发单分子测序技术来填补这一技术空白 能够直接识别四种典型核苷酸以及 I，所有这些都具有高分辨率和高 准确性。反过来，这样的技术发展将提高对 RNA 功能的理解，包括 I 的作用和意义，以实现更好的诊断、预后和治疗。在这最后 在第二阶段的努力中，EBS 将开发出 beta 原型 I 测序系统并展示其完整的 各种 RNA 类型的功能和分析能力以及测序基准 与当前最先进的方法相比的结果。