Exosome packaging of nucleic acids
AssigneeUniversity of Ottawa
InventorsDerrick Gibbings, James Andrew Taylor
Life of this patent
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Abstract
A method for preparing exosomes or exosome-like vesicles packaged with a nucleic acid of interest is provided. In certain embodiments, the method may comprise: introducing into an exosome-producing cell a nucleic acid construct comprising the nucleic acid sequence of interest incorporated in a pre-miR-451 structural mimic, and allowing the cell to produce exosomes. Nucleic acid constructs, compositions, and uses thereof are also provided.
Claims15
A method for producing exosomes or exosome-like vesicles comprising a gene silencing nucleic acid, a nucleic acid of interest, or a precursor thereof, said method comprising:
- introducing into an exosome-producing cell, or expressing in an exosome-producing cell, a nucleic acid construct comprising the gene silencing nucleic acid, nucleic acid of interest, or a precursor thereof, incorporated within a pre-miR-451 structural mimic;
- treating the exosome-producing cell with a lysosomal or autophagy inhibitor;
- producing exosomes or exosome-like vesicles from the cell;
- and
- collecting or enriching the produced exosomes or exosome-like vesicles.
A method for producing exosomes or exosome-like vesicles comprising a gene silencing nucleic acid, a nucleic acid of interest, or a precursor thereof, said method comprising:
- introducing into an exosome-producing cell, or expressing in an exosome-producing cell, a nucleic acid construct comprising the gene silencing nucleic acid, nucleic acid of interest, or a precursor thereof, incorporated within a pre-miR-451 structural mimic;
- producing exosomes or exosome-like vesicles from the cell;
- and
- collecting or enriching the produced exosomes or exosome-like vesicles,
- wherein the pre-miR-451 structural mimic comprises a stem-loop secondary structure having at least one base pair mismatch in the stem, and wherein at least one base pair mismatch in the stem is positioned within the first three base pairs adjacent to a Drosha cleavage site.
A method for producing exosomes or exosome-like vesicles comprising a gene silencing nucleic acid, a nucleic acid of interest, or a precursor thereof, said method comprising:
- introducing into an exosome-producing cell, or expressing in an exosome-producing cell, a nucleic acid construct comprising the gene silencing nucleic acid, nucleic acid of interest, or a precursor thereof, incorporated within a pre-miR-451 structural mimic;
- producing exosomes or exosome-like vesicles from the cell;
- and
- collecting or enriching the produced exosomes or exosome-like vesicles,
- wherein the pre-miR-451 structural mimic comprises a single-stranded structure including a 3′ portion, which is a loop-derived sequence, mimicking mature miR-451.
A method for producing exosomes or exosome-like vesicles comprising a gene silencing nucleic acid, a nucleic acid of interest, or a precursor thereof, said method comprising:
- introducing into an exosome-producing cell, or expressing in an exosome-producing cell, a nucleic acid construct comprising the gene silencing nucleic acid, nucleic acid of interest, or a precursor thereof, incorporated within a pre-miR-451 structural mimic;
- producing exosomes or exosome-like vesicles from the cell;
- and
- collecting or enriching the produced exosomes or exosome-like vesicles,
- wherein the pre-miR-451 structural mimic comprises a single-stranded structure including a 3′ portion, mimicking mature miR-451, and wherein the pre-miR-451 structural mimic is about 23-24 nt in length.
Dependencies are inferred from the claim text.