Detection of Methylation and Translocation Events by Novel Sequencing Technology

应用新型测序技术检测甲基化和易位事件 摘要 DNA测序技术的迅速发展，使我们对DNA序列的分子水平有了更深入的了解。 癌症的基础然而，癌症基因组学涉及非常复杂的分子重排，如 易位，需要调查长的DNA分子与单核苷酸歧视。电流 下一代测序（NGS）技术不能解决这样的性能要求，并且面临 挑战局限性。 这项提案的长期目标是开发一种快速和高度准确的DNA测序系统 它有可能完全控制DNA通过电子通道的速度和方向， 探测器（固态）在正向（阅读）和反向（校对）方向。与其他 纳米孔技术，我们的技术将进料步骤与阅读步骤分开， 从微量材料开始测序全基因组。结合起来，这些特征具有 为我们的测序技术提供实现单核苷酸区分的能力的潜力 使用长DNA分子（200kb）的准确度为99.9%或更高。预计这样的系统将 能够从微量的材料开始准确地阅读这样长的DNA片段， 扩大的需要。这将为从业者和癌症研究人员提供更好的 工具比目前最先进的测序技术。事实上，我们预计我们的技术将 能够收集表达和非表达的癌症的遗传和表观遗传标记的信息， 表达基因组的区域，同时达到目前尚未实现的时间和成本性能指标。 市场 当前第1阶段（R43）提案的目标是通过结合我们的专有技术 如果我们将这项技术应用到测序系统中，我们将能够检测到重要的分子变化， 与癌症发生和增殖相关，包括长距离甲基化和易位， 以及铬。由于我们的系统能够完全控制 通过测序系统检测器时DNA的易位率和线性化。 在第二阶段（R44）将集中在嵌入式固态（电子）纳米- 第一阶段开发的用于单碱基核苷酸阅读的初始系统的电极检测组件。 如果目标里程碑得以实现，则原型仪器的商业开发将沿着 与相关的耗材芯片和试剂将立即启动后，完成 第二阶段。 我们预计，我们的技术有可能在不到一个小时的时间内完成一个完整的人类基因组测序。 小时，成本为200美元的活检材料，从而达到性能指标目前无法在 目前的市场，并提供癌症研究人员和医疗从业者与一个宝贵的负担得起的工具 用于阐明癌症的分子机制和检测特定的分子变化， 可以改善疾病的早期临床检测。

Detection of Methylation and Translocation Events by Novel Sequencing Technology ABSTRACT The rapid development of DNA sequencing technologies advanced our understanding of the molecular basis of cancer. However, cancer genomics involves very complex molecular rearrangements, such as translocations, requiring the investigation of long DNA molecules with single nucleotide discrimination. Current Next Generation Sequencing (NGS) technologies cannot address such performance requirements and face challenging limitations. The long term goal of this proposal is to develop a rapid and highly accurate DNA sequencing system that has the potential to fully control the speed and orientation at which the DNA passes through an electronic detector (solid state) both in forward (reading) and reverse (proof read) directions. In contrast with other nanopore technologies, our technology decouples the feeding step from the reading step, enabling to sequence full genomes starting with minute quantity of material. Combined, the characteristics have the potential to provide our sequencing technology with the capability to achieve single nucleotide discrimination with an accuracy of 99.9% or higher using long DNA molecules (200kb). Such a system is anticipated to be capable of accurately reading such long fragments of DNA starting from minute quantities of material without the need of amplification. This would provide both practitioners and cancer researchers with a much better tool than the current state of the art sequencing technologies. Indeed we project that our technology will enable the collection of information of genetic and epigenetic markers of cancer both in expressed and non expressed areas of the genome while reaching time and cost performance metrics currently unachieved on the market. The goal of the current Phase 1 (R43) proposal is to demonstrate that by incorporating our proprietary technology into a sequencing system, we will be able to detect important molecular changes that are associated with cancer initiation and proliferation, to include long range methylations and translocations as well as chromoplexy. Such detection will be rendered possible due to the ability of our system to fully control the translocation rate and linearization of the DNA while passing through the sequencing system detector. In Phase II (R44) will focus on the incorporation of an embedded solid state (electronic) nano- electrode detection component to the initial system developed in phase I for single base nucleotide reading. Provided that targeted milestones are achieved, commercial development of a prototype instrument along with associated consumable chips and reagents will be initiated immediately following the completion of phase II. We project that our technology has the potential to sequence a full human genome in less than one hour at the cost of $200 from biopsy material, thus reaching performance metrics currently unavailable on the current market, and providing cancer researchers and medical practitioners with an invaluable affordable tool for the elucidation of molecular mechanisms of cancer and the detection of specific molecular changes which may improve early clinical detection of the disease.