PARALLEL SYNTHESIS OF RNA OLIGONUCLEOTIDE MICROARRAYS

RNA寡核苷酸微阵列的平行合成

• 批准号: 6350431
• 负责人: XIAOLIAN GAO
• 金额: $ 14.75万
• 依托单位: UNIVERSITY OF HOUSTON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-15 至 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6350431
• 关键词: DNA; RNA; RNA binding protein; antisense nucleic acid; combinatorial chemistry; genetic library; genetic markers; microarray technology; nuclear magnetic resonance spectroscopy; nucleic acid chemical synthesis; nucleic acid hybridization; nucleic acid sequence; oligonucleotides; oncogenes; oncoproteins; photochemistry; protein kinase C

The objective of this proposal is to develop a novel technology for parallel synthesis of RNA microarray on solid surfaces (RNA-chips). Although DNA-chips have emerged as a high efficient and comprehensive tool for cancer gene analysis, drug discovery and a variety of bioassays, a parallel development of RNA-chips has not been possible, primarily because there is no practical method that can achieve efficient and economic parallel synthesis of RNA microarrays. In this proposal, a method of RNA-chip synthesis using novel photochemical reactions in combination with conventional chemistry of RNA synthesis will be developed. The method uses photogenerated reagents to allow optical control of a single reaction step, such as deprotection of blocking groups, in a sequence of reactions. Using a microarray reactor and a programmable digital optical projector, parallel synthesis of thousands of diverse RNA sequences can be achieved with high efficiency and low costs. Our Phase I goals are to obtain prototype RNA oligonucleotide arrays using the proposed method of light directed parallel synthesis and demonstrate hybridization patterns of the RNA-chips synthesized. A novel combinatory strategy using our chip technology will be applied to screen hundreds of photoreaction conditions. New applications of the light controlled RNA-chip synthesis will be explored. Our Phase II goals are to produce next generation of high quality RNA-chips containing oligonucleotides composed of 20-40 residues and establish the applications of RNA cancer gene-chips and RNA library-chips in identification of RNA targets by antisense oligonucleotides and potential antitumor agents. RNA-chips provide unambiguous sequence information and are indispensable and powerful tool for direct investigation of RNA interactions with a variety of ligand molecules in cancer analysis and drug development and in the study of many human diseases. It is expected that in keeping pace with rapidly mounting information in RNA genomics, there will be an escalated effort in identifying RNA sequences as specific diagnose and therapeutic targets. Our proposed research is to provide a highly efficient, flexible and easily accessible technology to accelerate these discovery processes.

本研究的目的是开发一种新的技术，用于在固体表面上并行合成RNA微阵列（RNA芯片）。 尽管DNA芯片已经成为癌症基因分析、药物发现和各种生物测定的高效和全面的工具，但RNA芯片的平行开发一直是不可能的，主要是因为没有实用的方法可以实现RNA微阵列的高效和经济的平行合成。在这个提议中，将开发一种使用新颖的光化学反应与RNA合成的常规化学相结合的RNA芯片合成方法。 该方法使用光生试剂，以允许在一系列反应中对单个反应步骤进行光学控制，例如封闭基团的脱保护。 使用微阵列反应器和可编程数字光学投影仪，可以以高效率和低成本实现数千种不同RNA序列的并行合成。我们的第一阶段的目标是获得原型RNA寡核苷酸阵列，使用所提出的方法，光定向平行合成，并证明杂交模式的RNA芯片合成。 利用我们的芯片技术，一种新的组合策略将被应用于筛选数百个光反应条件。 将探索光控RNA芯片合成的新应用。 我们的第二阶段目标是生产下一代高质量的含有20-40个残基的寡核苷酸的RNA芯片，并建立RNA癌症基因芯片和RNA文库芯片在识别RNA靶点和潜在的抗肿瘤药物中的应用。RNA芯片提供明确的序列信息，是直接研究RNA与各种配体分子相互作用的不可或缺的强大工具，用于癌症分析和药物开发以及许多人类疾病的研究。 预计在保持与RNA基因组学中快速增加的信息的步伐中，将有一个升级的努力来鉴定RNA序列作为特异性诊断和治疗靶标。 我们提出的研究是提供一种高效，灵活和易于访问的技术来加速这些发现过程。