Small molecule-regulated gene silencing technologies

自从最近被发现以来，RNA干扰（RNAi）已经成为生物学家选择性抑制几乎任何他们选择的基因表达的有力工具。RNAi帮助研究人员发现了新的基因功能以及各种基因如何相互作用，这对于理解许多生物过程和疾病的分子基础至关重要。此外，许多学术和工业团体正在集中开发利用RNAi治疗和治愈疾病的技术。因此，RNAi有望成为一项重要的基础科学和应用技术。该项目的目标是开发新的技术来控制小分子药物在何时何地诱导RNAi。化学调节的RNAi能够研究对细胞存活或生长至关重要的基因，观察动态细胞功能，并提高治疗应用的安全性。然而，目前可用的技术需要复杂工程蛋白的共表达，这可能导致免疫原性并发症或毒性。首席研究员（PI）提出的策略是利用RNA与各种化合物特异性相互作用的能力来设计RNA，使其同时充当化学传感器和RNAi诱导剂。结果是遗传上小的、无蛋白质的RNA纳米机器，它能感知细胞中的特定信号分子，并决定是否触发RNAi。尽管RNAi已迅速成为许多学术和工业实验室的常规技术，但学生，特别是本科生，很少有机会学习和实践这一重要技术。该项目将允许加州大学戴维斯分校生物医学工程专业的学生通过高级设计课程获得RNAi的实践经验。学生将以小组为单位，设计新的化学调控RNAi系统，构建质粒载体在哺乳动物细胞系中表达，并在两个学期内研究其功能。通过这个项目，学生将在一个团队合作的环境中获得宝贵的RNAi和各种生物工程技术的经验。此外，研究小组每年夏天将通过加州大学戴维斯分校青年学者计划接待一到两名高中生参与实验室研究。

CBET-0755053YokobaysahiIntellectual merit Since its recent discovery, RNA interference (RNAi) has emerged as a powerful tool for biologists to selectively repress expression of virtually any gene of their choice. RNAi has helped researchers to discover novel gene functions and how various genes interact, which are crucial to understanding the molecular basis of many biological processes and diseases. Additionally, numerous academic and industrial groups are intensively developing technologies to use RNAi to treat and cure diseases. Consequently, RNAi is expected to become an important technology for both basic science and applications. The goal of this project is to develop novel technologies to control when and where RNAi is induced by small molecule drugs. Chemically-regulated RNAi enables study of genes that are essential for cell survival or growth, observation of dynamic cellular functions, and increased safety for therapeutic applications. The presently available techniques, however, require coexpression of complex engineered protein(s), which can cause immunogenic complications or toxicity. The Principal Investigator's (PI's) proposed strategies harness the ability of RNA to specifically interact with a variety of compounds to engineer RNA to act as both a chemical sensor and RNAi inducer. The results are genetically small, protein-free, RNA nanomachines which sense specific signal molecules in the cell and decide whether or not to trigger RNAi. Broader impacts Although RNAi has quickly become a routine technique in many academic and industrial laboratories, students, especially undergraduate students, have very little opportunity to learn and practice the important technology. This project will allow a number of Biomedical Engineering students at UC Davis to gain hands-on experience with RNAi through the Senior Design course. The students will, in small groups, design novel chemically regulated RNAi systems, construct plasmid vectors to express them in mammalian cell lines, and study their functions over two academic quarters. Through this project, the students will gain valuable experience with RNAi and various bioengineering techniques in a team-oriented environment. Additionally, the research group will host one or two high school students per summer to engage in laboratory research through UC Davis Young Scholars Program.