Two readers for RNA

项目摘要 基于纳米孔的技术有可能成为全面RNA测序的未来，即， 转录组学和表转录组学，因为它是直接表征和分析长 没有逆转录或加工酶的RNA链不能穿过某些序列。的 通过纳米孔测序实现的直接表征也将最终导致检测， 修饰的碱基的表征，这是表转录组学的关键要求。目前可用的，国家- 现有技术的RNA测序技术受到该方法的限制，其输入需要cDNA中间体 和/或大量昂贵的样品，并且读出特征在于短读取长度和 低精度。在这个项目中，电子生物科学（EBS）的目标是开发一个全新的长读， 基于纳米孔的极高准确度测序技术， 测序，能够进行固有的序列交叉验证和扩展的灵敏度， 通用RNA测序。在第一阶段项目中，我们将建立一个全新的系统， 评估和优化相关的工作流程/方法，以测序规范核苷酸和特定的 临床相关的RNA修饰，并证明初始测序。在这个项目结束时，我们 将成功展示和开发一个全新的测序系统原型， 克服了限制当前RNA测序技术的已知挑战（包括其他纳米孔， 基于测序的方法）。由此产生的技术发展将是一个无标签，长读（> kbase）， 电子读出方法，能够提供一个新的和详细的了解RNA及其调控。

Project Summary Nanopore-based technology has the potential to be the future of comprehensive RNA sequencing, i.e., transcriptomics and epitranscriptomics, as it is the only viable path to directly characterizing and profiling long RNA strands without reverse transcription or processive enzymes that fail to traverse certain sequences. The direct characterization enabled by nanopore sequencing will also ultimately lead to detection and characterization of modified bases, a critical requirement for epitranscriptomics. The presently available, state- of-the art RNA sequencing technologies are limited by the method, with the input requiring cDNA intermediates and/or large amounts of costly sample, and the readout featuring a compromise between short read length and low-accuracy. During this program, Electronic BioSciences (EBS) aims to develop a completely new long-read, extremely high-accuracy nanopore-based sequencing technology that will enable de novo epitranscriptomic sequencing, capable of inherent sequence cross-validation and expanded sensitivity for more accurate and versatile RNA sequencing. During this Phase I project, we will build and multiplex an entirely new system, fully assess and optimize the associated workflow/methodology to sequence canonical nucleotides and specific clinically relevant RNA modifications, and demonstrate initial sequencing. At the conclusion of this project, we will have successfully demonstrated and developed an entirely new sequencing system prototype that overcomes the known challenges limiting current RNA sequencing technologies (including other nanopore- based sequencing approaches). The resulting technology development will be a label-free, long-read (>kbases), electronic readout method, capable of providing a new and detailed understanding of RNA and its regulation.