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US 11,998,635 B2Granted
Targeted extracellular vesicles comprising membrane proteins with engineered glycosylation sites
AssigneeNorthwestern University
InventorsJoshua N. Leonard, Michelle E. Hung
Life of this patent
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Abstract
Disclosed are extracellular vesicles comprising an engineered targeting protein for targeting the extracellular vesicles to target cells. The targeting protein is a fusion protein that includes a ligand, an engineered glycosylation site, and an exosome-targeting domain. Exemplary extracellular vesicles may include but are not limited to exosomes.
Claims18
A method of loading an extracellular vesicle with a cargo nucleic acid, the method comprising the steps of:
- expressing within a vesicle-producing cell that contains the cargo nucleic acid (i)
- a fusion protein comprising:
- (i) a nucleic acid-binding domain;
- (ii) a vesicle-targeting domain, and
- (iii) an engineered glycosylation site, wherein the engineered glycosylation site comprises an amino acid sequence that is not naturally present in the fusion protein or any of the components of the fusion protein;
- wherein:
- the cargo nucleic acid includes a binding motif that binds to the nucleic acid-binding domain of the fusion protein, and
- the vesicle-targeting domain of the fusion protein localizes to the extracellular vesicles released by the vesicle-producing cell,
- such that the cargo nucleic acid is loaded into the vesicle when the fusion protein and cargo nucleic acid are both present in by the vesicle-producing cell.
6 more elements
In a method of loading an extracellular vesicle with a cargo nucleic acid, the method comprising the steps of:
- expressing within a vesicle-producing cell that contains the cargo nucleic acid
- a fusion protein comprising:
- (i) a nucleic acid-binding domain;
- and
- (ii) a vesicle-targeting domain, and
- wherein:
- the cargo nucleic acid includes a binding motif that binds to the nucleic acid-binding domain of the fusion protein, and
- the vesicle-targeting domain of the fusion protein localizes the extracellular vesicles released by the vesicle-producing cell,
- such that the cargo nucleic acid is loaded into the vesicle when the fusion protein and cargo nucleic acid are both present in the vesicle-producing cell;
- the improvement that comprises engineering the fusion protein to comprise an engineered glycosylation site, wherein the engineered glycosylation site comprises an amino acid sequence that is not naturally present in the fusion protein or any of the components of the fusion protein.
7 more elements
Dependencies are inferred from the claim text.