Department of Molecular Microbiology & Immunology, University of Missouri, Columbia, MO, 65212, USA.
Bond Life Sciences Center, University of Missouri, Columbia, MO, 65211, USA.
Nat Commun. 2018 Jun 11;9(1):2283. doi: 10.1038/s41467-018-04691-x.
Large RNAs and ribonucleoprotein complexes have powerful therapeutic potential, but effective cell-targeted delivery tools are limited. Aptamers that internalize into target cells can deliver siRNAs (<15 kDa, 19-21 nt/strand). We demonstrate a modular nanostructure for cellular delivery of large, functional RNA payloads (50-80 kDa, 175-250 nt) by aptamers that recognize multiple human B cell cancer lines and transferrin receptor-expressing cells. Fluorogenic RNA reporter payloads enable accelerated testing of platform designs and rapid evaluation of assembly and internalization. Modularity is demonstrated by swapping in different targeting and payload aptamers. Both modules internalize into leukemic B cell lines and remained colocalized within endosomes. Fluorescence from internalized RNA persists for ≥2 h, suggesting a sizable window for aptamer payloads to exert influence upon targeted cells. This demonstration of aptamer-mediated, cell-internalizing delivery of large RNAs with retention of functional structure raises the possibility of manipulating endosomes and cells by delivering large aptamers and regulatory RNAs.
大型 RNA 和核糖核蛋白复合物具有强大的治疗潜力，但有效的细胞靶向输送工具有限。能够内化进入靶细胞的适体可以递送 siRNA（<15 kDa，19-21 nt/链）。我们通过识别多种人类 B 细胞癌细胞系和转铁蛋白受体表达细胞的适体，展示了一种用于细胞内递大型功能性 RNA 有效载荷（50-80 kDa，175-250 nt）的模块化纳米结构。通过荧光报告 RNA 有效载荷，可以加速平台设计的测试，并快速评估组装和内化。通过替换不同的靶向和有效载荷适体来展示模块性。这两个模块都能内化进入白血病 B 细胞系，并在内涵体中保持共定位。内化的 RNA 发出的荧光持续至少 2 小时，这表明适体有效载荷有足够的时间对靶细胞施加影响。本研究证明了通过适体介导的、细胞内化的方式递大型 RNA 并保持其功能结构的保留，这为通过递大型适体和调节 RNA 来操纵内涵体和细胞提供了可能性。
