Journal
MOLECULAR THERAPY
Volume 25, Issue 7, Pages 1580-1587Publisher
CELL PRESS
DOI: 10.1016/j.ymthe.2017.03.021
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Funding
- Estonian Research Council Personal Research Grant and European Union FP7
- Innovative Medicines Initiative consortium COMPACT [115363]
- VENI Fellowship from the Netherlands Organisation for Scientific Research (NWO)
- BBSRC [BB/M024393/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/M024393/1] Funding Source: researchfish
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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 degrees C for 1 hr in 100 p.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.
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