Selectively expressed RNA molecules as a versatile tool for functionalized cell targeting. Selectively expressed RNA molecules as a versatile tool for functionalized cell targeting

Exact targeting of specific mammalian cell types or diseased cells is one of the most urgently needed prerequisites for a new generation of potent pharmaceuticals. Different approaches have been pursued, failing mainly due to a lack of specific surface markers in most cases. Developing a novel RNA-based methodology, we can now ensure exact cell targeting and simultaneously combine this with selective expression of effector proteins, thereby functionalization of the target cell for therapy, diagnostics or cell steering. The specific combination of the molecular properties of antisense technology and mRNA therapy with functional RNA secondary structures based on Nano-DMS-MaP analysis allowed us to develop selectively expressed RNA molecules for medical applications. These so-called seRNAs remain inactive in non-target cells and are only activated by partial degradation to induce translation in preselected cell types of interest. Cell type specificity and type of functionalization are easily adaptable based on a simple modular system. In proof of concept in vitro and in vivo studies we used seRNAs as a highly selective platform technology for powerful cell targeting. We effectively treat breast tumor cell clusters in mixed cell systems and reduce early brain tumors of mice without detectable side effects with just a single treatment. Our data open up new potential avenues for various therapeutic applications. Overall design: Nano-DMS-MaP analysis of seRNA. Selective and non-selective constructs we transfected into cells. RNA structure was determined by Nano-DMS-MaP probing.

精准靶向特定哺乳动物细胞类型或病变细胞，是新一代强效药物研发最为迫切的核心前提之一。目前学界已探索多种研发路径，但多数因缺乏特异性表面标志物而宣告失败。
本研究开发新型基于核糖核酸（RNA）的技术方法，如今可实现精准细胞靶向，并可同时结合效应蛋白的选择性表达，从而实现靶细胞的功能化，用于治疗、诊断或细胞调控。
基于Nano-DMS-MaP分析得到的功能性RNA二级结构，结合反义技术与信使RNA（mRNA）疗法的分子特性，我们得以开发可用于医学应用的选择性表达RNA分子（seRNAs，selectively expressed RNA）。
这类seRNAs在非靶细胞中保持无活性状态，仅通过部分降解被激活，从而在预先选定的目标细胞类型中诱导翻译过程。
基于简易模块化系统，细胞类型特异性与功能化类型均可便捷调整。
在体外与体内概念验证研究中，我们将seRNAs用作实现强效细胞靶向的高选择性平台技术。
仅需单次给药，即可在混合细胞体系中有效治疗乳腺肿瘤细胞团，并缩小小鼠早期脑肿瘤，且未检测到明显副作用。
本研究数据为各类治疗应用开辟了全新的潜在研发路径。
整体实验设计：对seRNA开展Nano-DMS-MaP分析。将选择性与非选择性构建体转染至细胞中，通过Nano-DMS-MaP探针法测定RNA二级结构。