RNA

Part:BBa_K1633004:Experience

Designed by: Chen Xi, Zhou Yu, Jiang Waner, Zhang Peng, Tian Chenfei   Group: iGEM15_NJU-China   (2015-09-13)
Revision as of 11:36, 18 September 2015 by ChenXi (Talk | contribs) (→‎Applications of BBa_K1633004)


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Applications of BBa_K1633004

USAGE AND BIOLOGY

We package MOR siRNA into exosomes by transfecting HEK293 cells with a a Lamp2b-RVG plasmid and the MOR siRNA-2 plasmid and then collect siRNA-encapsulated exosomes. When inject the modified exosomes into the bloodstream, exosome will specifically recognize acetylcholine receptors and fuse with neurons under the direction of the RVG peptide. Once inside neurons, MOR siRNA will degrade MOR mRNA by base-pairing, resulting in sharp decrease of MOR on neuron membrane. As a consequence, MOR reduction and disturbed function will result in the inhabitation of the secretion of GABA and the suppression of the dopaminergic reward pathway, which ultimately have some therapeutic effects on opioid dependence.

CHARACTERIZATION

Interference efficiency of MOR siRNA-2 plasmid

To ensure the interference efficiency,MOR siRNA-2 plasmid was transfected into the mouse neuroblastoma cell line Neuro2A. Efficient knockdown of MOR by MOR siRNA-2 in Neuro2A cells is observed.

NJU-China-parts-fig 4.png

Package of MOR siRNA into exosomes

The levels of MOR siRNA in isolated exosomes were assayed by a quantitative RT-PCR assay. The MOR siRNA concentration in exosomes was calculated to be approximately 0.14 pmol/μg. The results showed that MOR siRNA can be successfully packaged into exosomes, no matter the exosomes were modified on the outside membrane with or without RVG peptide.

NJU-China-parts-fig 5.png

User Reviews

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