Exploring the RNA syntax for engineering therapeutic RNA-based nano-factories

描述（由申请人提供）：小干扰sirna、核酶和反义rna等治疗剂在新的分子方法中显示出显著的潜力，可以下调癌细胞或病毒感染细胞中的特定基因表达。因此，开发安全、高效、特异性和非致病性的纳米颗粒来包装和递送多功能治疗性rna是非常需要的。虽然可编程RNA自组装已被证明有可能为实现这一目标提供解决方案，但基于RNA的多功能纳米结构的设计和工程的基本原理仍有待阐明。本提案的长期目标是促进基于RNA和核酸类似物的完全可控和多功能生物材料的发展，用于生物学研究，疾病诊断或治疗。为了实现这一挑战，该项目旨在生成刺激响应的可编程三维（3D）超分子组件，这些组件可以控制其几何形状、拓扑结构、方向性和可寻址性，并能够响应环境的提示。例如，可编程的、可寻址的和多功能的3D纳米笼，允许在3D空间中精确固定各种具有催化或识别特性的功能治疗模块，将被合成并通过生化和生物物理方法（如AFM和冷冻电子显微镜）进行表征。其中一些RNA纳米装置将被设计成在不同的超分子形状之间可逆地切换，以响应小的目标化合物。将研究协同和瞬态、时间激活的复杂功能的工程原理，以及使用RNA结构模拟物（如锁定核酸（LNA））增强这些组装的化学稳定性。这项工作将为控制涉及大量RNA分子的自组装过程提供新的见解，并将为设计具有强大生物学和医学潜力的复杂RNA纳米机器铺平道路。预计该项目将为设计和开发治疗性rna的多功能反应载体建立基本的组装和功能原则，这些载体将适用于体内应用并适应未来的工业规模发展。

DESCRIPTION (provided by applicant): Therapeutic agents such as small interfering siRNAs, ribozymes, and anti-sense RNAs show significant potential in new molecular approaches to down-regulate specific gene expression in cancerous or viral- infected cells. The development of safe, efficient, specific and non-pathogenic nano-particles for the packaging and delivery of multifunctional therapeutic RNAs is thus highly desirable. While programmable RNA self-assembly has been shown to potentially provide a solution towards that goal, the underlying principles for the design and engineering of multifunctional nano-structures based on RNA have still to be delineated. The long-term goal of this proposal is to contribute to the development of fully controllable and versatile bio-materials based on RNA and nucleic acid analogs for use in the study of biology, disease diagnosis, or therapy. In order to achieve this challenge, this project aims at generating stimuli-responsive programmable three-dimensional (3D) supra-molecular assemblies with control over their geometry, topology, directionality and addressability and able to respond to the cues of the environment. For instance, programmable, addressable and multifunctional 3D nano-cages that allow precise immobilization in 3D space of various functional therapeutic modules with catalytic or recognition properties will be synthesized and characterized by biochemical and biophysical methods such as AFM and cryo electron microscopy. Some of these RNA nano-devices will be engineered to switch reversibly between distinct supra-molecular shapes in response to small target compounds. The principles underlying the engineering of concerted and transient, time-activated complex functions as well as the enhancement of the chemical stability of these assemblies using RNA structural mimics like locked-in nucleic acid (LNA) will be investigated. This work will provide new insights in the control of self-assembly processes involving large population of RNA molecules and will pave the way towards the design of complex RNA-based nano-machines with strong potential in biology and medicine. It is anticipated that this project will establish the fundamental assembly and functional principles for the design and development of versatile responsive vehicles for therapeutic RNAs that will be suitable for in vivo applications and amenable to industrial-scale development in the future.