Nanoscale Tools for Inosine Sequencing

项目摘要 在此期间，电子生物科学公司（EBS）将充分开发和展示一种全新的RNA 测序技术能够直接测序RNA编辑的最常见产物：肌苷（I）。 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）至I，即，A-to-I编辑，其中编辑的碱基在mRNA中编码不同， mRNA剪接，并使miRNA和tRNA在细胞中发挥功能。目前无法直接测序 具有定量读出的A至IRNA编辑位点（即，在给定的范围内分析或确定它们的比例 序列）严重限制了研究人员充分理解这种表转录组学过程的能力， 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 能够直接鉴定四个典型核苷酸沿着I，所有这些都具有高分辨率和高灵敏度， 精度反过来，这样的技术发展将提高对RNA功能的理解，包括 I的作用和重要性，以实现更好的诊断、诊断和治疗。于年底 在第二阶段的努力中，EBS将开发出一个beta原型I测序系统，并展示其完整的 功能和分析能力的各种RNA类型，以及基准的测序 结果与当前最先进的方法进行比较。

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.