用化学稳定的疏水性小干扰RNA装载细胞外囊泡用于治疗中枢神经系统疾病
Loading of Extracellular Vesicles with Chemically Stabilized Hydrophobic siRNAs for the Treatment of Disease in the Central Nervous System.
作者信息
Haraszti Reka A, Coles Andrew, Aronin Neil, Khvorova Anastasia, Didiot Marie-Cécile
机构信息
RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA, USA.
Department of Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA.
出版信息
Bio Protoc. 2017 Jun 20;7(20). doi: 10.21769/BioProtoc.2338.
Efficient delivery of oligonucleotide therapeutics, siRNAs, to the central nervous system represents a significant barrier to their clinical advancement for the treatment of neurological disorders. Small, endogenous extracellular vesicles were shown to be able to transport lipids, proteins and RNA between cells, including neurons. This natural trafficking ability gives extracellular vesicles the potential to be used as delivery vehicles for oligonucleotides, siRNAs. However, robust and scalable methods for loading of extracellular vesicles with oligonucleotide cargo are lacking. We describe a detailed protocol for the loading of hydrophobically modified siRNAs into extracellular vesicles upon simple co-incubation. We detail methods of the workflow from purification of extracellular vesicles to data analysis. This method may advance extracellular vesicles-based therapies for the treatment of a broad range of neurological disorders.
将寡核苷酸疗法、小干扰RNA（siRNAs）有效递送至中枢神经系统是其在治疗神经疾病方面临床进展的重大障碍。小型内源性细胞外囊泡已被证明能够在包括神经元在内的细胞之间运输脂质、蛋白质和RNA。这种天然的运输能力使细胞外囊泡有潜力用作寡核苷酸、siRNAs的递送载体。然而，目前缺乏将寡核苷酸货物加载到细胞外囊泡中的强大且可扩展的方法。我们描述了一种简单共孵育后将疏水修饰的siRNAs加载到细胞外囊泡中的详细方案。我们详细介绍了从细胞外囊泡纯化到数据分析的工作流程方法。该方法可能会推动基于细胞外囊泡的疗法用于治疗广泛的神经疾病。
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