Integrated RNA Amplification Micro Card

集成RNA扩增微卡

• 批准号: 7479999
• 负责人: Jason A.A. West
• 金额: $ 24.21万
• 依托单位: ARCXIS BIOTECHNOLOGIES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7479999
• 关键词: Achievement; Address; Biochemical Process; Biochemical Reaction; Biological; Biology; Biomedical Research; Biotechnology; Cell physiology; Classification; Clinical; Companions; Complement component C1s; Complementary DNA; Complex; DNA Polymerase I; Devices; Diagnostic; Diagnostic Procedure; Diagnostics Research; Disease; Disease Progression; Evaluation; Evolution; Foundations; Future; Gene Expression Profiling; Genetic Materials; Genetic Transcription; Goals; Government; Heating; Human; Label; Laboratories; Liquid substance; Manuals; Messenger RNA; Methods; Microarray Analysis; Monitor; Nucleic Acids; Operative Surgical Procedures; Output; Performance; Persons; Phase; Polymerase; Polymerase Chain Reaction; Polymers; Positioning Attribute; Preparation; Procedures; Process; Productivity; Pump; Purpose; RNA; RNA amplification; RNA chemical synthesis; RNA purification; RNA-Directed DNA Polymerase; Rate; Reaction; Reagent; Research; Resources; Reverse Transcription; Ribonuclease H; Route; Running; Sampling; Silicon Dioxide; Surface; System; Techniques; Technology; Temperature; Time; Today; base; concept; cost; cost effective; design; experience; fluid flow; genetic analysis; instrument; instrumentation; internal control; mRNA Expression; novel; oligo (dT); pressure; prototype; research study; response; tool; user-friendly

DESCRIPTION (provided by applicant): Nucleic acid analysis is the essential foundation of many biomedical research and diagnostic methods. Among these, microarray-based gene expression profiling has become a powerful research tool for understanding basic disease biology, and a powerful diagnostic tool for profiling cellular states during disease progression and in response to treatment. However, preparation of RNA from biological samples for microarray analysis is presently a rate-limiting bottleneck. This is because RNA purification, amplification, and labeling remains a complex, time-consuming, labor-intensive, and expensive process. The primary goal of this proposal is produce devices and systems that dramatically reduce this burden, so that it is no longer an impediment to health-related research and diagnostics. A second goal is to increase experimental reliability, by reducing human error, thereby increasing the utility and efficiency of microarray expression analysis methods. To accomplish these goals Arcxis Biotechnolgies will develop a microamplification card (MAC), a small, integrated fluidic processing device, and a companion bench-top instrument that will ultimately completely automate sample preparation for microarray expression analysis. Multiple cards will run simultaneously in the MAC instrument, which will occupy only a small amount of laboratory bench space, greatly increasing laboratory productivity and decreasing utilization of laboratory resources. The initial goal of the proposed research is to demonstrate integration of linear mRNA amplification and amplified RNA purification on a microamplification card. The specific aims of the proposed research involve, first, fabrication of microamplification cards and actuating instrumentation, and second, systematic implementation and characterization of the biochemical reactions involved in linear RNA amplification and RNA purification. Both of these biochemical processes are well-established and highly characterized, allowing research to focus on their specific implementation in microamplification cards. Integration of biochemical processes of this complexity in a commercially viable format will constitute a significant achievement. First, it paves the way for rapid integration of additional processes that will enable end-to-end processing, from cell lysates to pure, amplified, labeled mRNA ready for microarray analysis. Second, it will provide a platform fluidic technology that will find wider application in biomedical research in the future. Microarray-based gene expression profiling has become a powerful research tool for understanding basic disease biology, and a powerful diagnostic tool for profiling cellular states during disease progression and in response to treatment. However, RNA purification, amplification, and labeling for microarrays, remain complex, time-consuming, labor-intensive, and expensive processes. The primary goal of this proposal is produce devices and systems that dramatically reduce this burden, so that it is no longer an impediment to health-related research and diagnostics.

描述（由申请人提供）：核酸分析是许多生物医学研究和诊断方法的基础。其中，基于微阵列的基因表达分析已成为理解基本疾病生物学的强大研究工具，并且是在疾病进展过程中和治疗响应过程中分析细胞状态的强大诊断工具。但是，从生物样品中制备用于微阵列分析的RNA目前是限制瓶装的瓶颈。这是因为RNA纯化，放大和标记仍然是一个复杂，耗时，劳动力且昂贵的过程。该提案的主要目标是生产大大减轻这种负担的设备和系统，因此不再是与健康相关的研究和诊断的障碍。第二个目标是通过减少人体错误来提高实验可靠性，从而增加微阵列表达分析方法的效用和效率。为了实现这些目标，Arcxis Biotechnolgies将开发微型放大卡（MAC），一个小型的集成流体处理设备，以及一种伴随的台式仪器，最终将完全自动化微阵列表达分析的样品制备。多张卡将同时在MAC仪器中运行，这将仅占据少量的实验室长凳空间，从而大大提高了实验室生产率并降低了实验室资源的利用率。拟议的研究的最初目标是证明在微扩增卡上的线性mRNA扩增和放大RNA纯化的整合。拟议的研究的具体目的是首先是制造微型放大卡和致动仪器，其次是系统性RNA扩增和RNA纯化涉及的生化反应的系统实施和表征。这两个生化过程都是完善的，并且具有很高的特征，从而使研究专注于它们在微型放大卡中的特定实现。以商业上可行的格式整合这种复杂性的生化过程将构成一个重大成就。首先，它为快速整合的其他过程铺平了道路，这些过程将使从细胞裂解物到纯，放大，标记的mRNA可以进行微阵列分析。其次，它将提供一种平台流体技术，将来会在生物医学研究中找到更广泛的应用。基于微阵列的基因表达分析已成为理解基本疾病生物学的强大研究工具，并且是在疾病进展过程中和对治疗响应过程中分析细胞状态的强大诊断工具。但是，用于微阵列的RNA净化，扩增和标记，保持复杂，耗时，劳动密集型和昂贵的过程。该提案的主要目标是生产大大减轻这种负担的设备和系统，因此不再是与健康相关的研究和诊断的障碍。