siRNA Encapsulation into Exosomes Using Electroporation: An Electric Pulse Based Technique to Load siRNA into Exosomes
Published: April 30, 2023
Abstract
Source: Faruqu, F. N. et al. Preparation of Exosomes for siRNA Delivery to Cancer Cells. J. Vis. Exp. (2018)
This video describes an electroporation-based method for encapsulating siRNA into exosomes. The siRNA-loaded exosomes can then be used for therapeutic applications.
Protocol
1. siRNA Encapsulation into Exosomes by Electroporation Pre-chill the electroporation cuvette (see Table of Materials) on ice for 30 min before electroporation. Mix 7.0 µg of exosomes (32 µL from 7 x 1012 p/mL stock in PBS) with 0.33 µg of siRNA (12 µL from 2 µM stock in RNase-free water) in the microcentrifuge tube. Make up the volume to 150 µL with citric acid buffer (see Table of Materials). The exosome to…
Divulgations
The authors have nothing to disclose.
Materials
| Atto655-siRNA | Eurogentec | SQ-SIRNA | (Labelled-S) UGC-GCU-ACGAUC-GAC-GAU-G55; (UnlabelledAS) CAU-CGU-CGA-UCG-UAGCGC-A55. |
| Glass pipettes | Fisher Scientific | 1156-6963 | |
| Amaxa Nucleofector I | Lonza | Nucleofector I | With Amaxa’s Nucleofector kits |
| Citric acid buffer with EDTA | Mix 0.1954 g citric acid and 0.2087 g disodium phosphate in 50 mL of deionised water. Add EDTA to 0.1 mM. Adjust pH to 4.4. | ||
| Microfuge tubes | Starlab | S1615-5500 | 1.5 ml |
Tags
Citer Cet Article
siRNA Encapsulation into Exosomes Using Electroporation: An Electric Pulse Based Technique to Load siRNA into Exosomes. J. Vis. Exp. (Pending Publication), e20566, doi: (2023).