Difference between revisions of "Part:BBa K2976015"

Revision as of 18:11, 21 October 2019

targeting exosome-NF-κB induced hsa-let-7f

targeting exosome-NF-κB induced hsa-let-7f

Usage

LAMP2, highly expressed on exosome membrane and linked with PD-L1-targeting peptide by (Gly4Ser)3, could not endow exosomes with macrophage-targeting ability. What´s more, the glycosylation protection help avoid excision of signal peptides and increase their expression on exosomes. With the utilization of such targeting exosomes, miRNA hsa-let-7f delivered could be increased in infected macrophages and the treatment of intracellular Mtb could be realized. HA tag is inserted into the fusion proteins on the membrane of exosomes for the immunoblotting assay.

Biology

MicroRNA hsa-let-7f is reported to regulate human immune responses to Mtb via control of A20, an inhibitor of the NF-κB pathway, which plays an essential role in regulating the expression of pro-inflammatory cytokine and chemokine genes. However, when Mtb infects macrophages, hsa-let-7f inside macrophages decreases. So exogenous supplementation of let-7f increases the level of endogenous let-7f and further up regulates NF-κB, thus can boost the secretion of cytokines, chemokines and NO to kill intracellular Mtb. To improve the targeting effectiveness of hsa-let-7f contained exosome , anti-PD-L1 peptide is displayed on the surface of the exosome to specifically target to the Mtb-infected macrophages. At the same time, Lamp 2, a protein which is highly expressed on the surface of exosomes assists anti-PD-L1 peptide to find Mtb-infected macrophages.

Characterization

Sequence and Features