Enzyme Bioreactor for Optimized Production of Modified Nucleic Acids

描述（由申请人提供）：我们将开发优化的酶法生产改性寡核苷酸的方法，这些方法作为治疗剂和生物传感器具有很强的优势。我们将首先实现具有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。我们将利用该平台优化2'- o -甲基和2'-氟RNA的体外转录条件。我们还将实施一个连续、连续或半连续流动的生物反应器，用于体外生产修饰RNA，用于正在进行的项目，包括微流控癌细胞捕获和适体治疗的体内评估。我们将建造的这种微反应器已被证明可用于控制、高效、可扩展的合成有机化学。这些相同的优点使该策略对生化反应具有吸引力。

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.