Enzyme Bioreactor for Optimized Production of Modified Nucleic Acids

用于优化生产修饰核酸的酶生物反应器

• 批准号: 8003777
• 负责人: Peter B Allen
• 金额: $ 4.76万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8003777
• 关键词: Biochemical; Biochemical Reaction; Bioreactors; Biosensor; Blood; Chemicals; DNA; Data Collection; Environment; Enzymes; Evaluation; Genetic Transcription; In Vitro; Injection of therapeutic agent; Methods; Microfluidics; Nucleic Acids; Oligonucleotides; Organic Chemistry; Pharmaceutical Preparations; Production; Protocols documentation; RNA; Resistance; Therapeutic; Therapeutic Agents; aptamer; cancer cell; improved; in vivo; instrument; public health relevance; research clinical testing; tool

DESCRIPTION (provided by applicant): We will develop optimized methods for the enzymatic production of modified oligonucleotides which have strong advantages as therapeutic agents and biosensors. We will first implement a Sequential Injection Analysis (SIA) platform with continuous data collection to optimize biochemical protocols with large parameter spaces. We will use this platform to optimize conditions for in vitro transcription of 2'-O-methyl and 2'-fluoro RNA. We will also implement a continuous, continuous- or semi-continuous-flow bioreactor for in vitro production of modified RNA for use in ongoing projects including microfluidic cancer cell capture and in vivo evaluations of aptamer therapeutics. Microreactors of the type we will build have been demonstrated for controlled, efficient, scalable, synthetic organic chemistry. These same advantages make the strategy attractive for biochemical reactions. PUBLIC HEALTH RELEVANCE: Chemically modified DNA and RNA are more resistant to being destroyed by the environment in blood, and so they are preferred as tools to develop drugs or instruments used for clinical testing. Current methods for generating these valuable substances are very expensive and produce very little of the product, which limits their use. This project will develop an improved method to generate modified DNA and RNA which will allow wider use of these important chemical tools.

描述（由申请人提供）：我们将开发用于酶促生产修饰的寡核苷酸的优化方法，所述修饰的寡核苷酸作为治疗剂和生物传感器具有很强的优势。我们将首先实施连续数据收集的顺序注射分析（SIA）平台，以优化具有大参数空间的生化方案。我们将使用这个平台来优化2 '-O-甲基和2'-氟RNA的体外转录条件。我们还将实施连续、连续或半连续流动生物反应器，用于体外生产修饰的RNA，用于正在进行的项目，包括微流体癌细胞捕获和适体治疗的体内评估。我们将建造的这种微型反应器已经被证明是可控的、高效的、可扩展的、合成有机化学。这些相同的优点使得该策略对于生化反应具有吸引力。 公共卫生相关性：化学修饰的DNA和RNA更能抵抗血液中环境的破坏，因此它们被首选为开发用于临床测试的药物或仪器的工具。目前用于产生这些有价值物质的方法非常昂贵并且产生非常少的产品，这限制了它们的使用。该项目将开发一种改进的方法来产生修饰的DNA和RNA，这将允许更广泛地使用这些重要的化学工具。