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Functional Delivery of Lipid-Conjugated siRNA by Extracellular Vesicles.


ABSTRACT: Extracellular vesicles (EVs) are cell-derived, membranous nanoparticles that mediate intercellular communication by transferring biomolecules, including proteins and RNA, between cells. As a result of their suggested natural capability to functionally deliver RNA, EVs may be harnessed as therapeutic RNA carriers. One major limitation for their translation to therapeutic use is the lack of an efficient, robust, and scalable method to load EVs with RNA molecules of interest. Here, we evaluated and optimized methods to load EVs with cholesterol-conjugated small interfering RNAs (cc-siRNAs) by systematic evaluation of the influence of key parameters, including incubation time, volume, temperature, and EV:cc-siRNA ratio. EV loading under conditions that resulted in the highest siRNA retention percentage, incubating 15 molecules of cc-siRNA per EV at 37°C for 1 hr in 100 ?L, facilitated concentration-dependent silencing of human antigen R (HuR), a therapeutic target in cancer, in EV-treated cells. These results may accelerate the development of EV-based therapeutics.

SUBMITTER: O'Loughlin AJ 

PROVIDER: S-EPMC5498810 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature

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Functional Delivery of Lipid-Conjugated siRNA by Extracellular Vesicles.

O'Loughlin Aisling J AJ   Mäger Imre I   de Jong Olivier G OG   Varela Miguel A MA   Schiffelers Raymond M RM   El Andaloussi Samir S   Wood Matthew J A MJA   Vader Pieter P  

Molecular therapy : the journal of the American Society of Gene Therapy 20170406 7


Extracellular vesicles (EVs) are cell-derived, membranous nanoparticles that mediate intercellular communication by transferring biomolecules, including proteins and RNA, between cells. As a result of their suggested natural capability to functionally deliver RNA, EVs may be harnessed as therapeutic RNA carriers. One major limitation for their translation to therapeutic use is the lack of an efficient, robust, and scalable method to load EVs with RNA molecules of interest. Here, we evaluated and  ...[more]

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