Delivery of therapeutic agent
A method of producing nanovesicles comprising an oligonucleotide inhibitor to an oncogene or a proto-oncogene or the gene product thereof, said method comprises a) introducing a DNA sequence encoding an oligonucleotide capable of inhibiting a human oncogenic or proto-oncogenic transcription factor, into a mammalian cell; b) allowing the cell to express said inhibitor oligonucleotide; and c) obtaining nanovesicles containing said inhibitor oligonucleotide from said cell. Nanovesicles produced by the claimed method can be effectively and specifically targeted to e.g. cancer cells to deliver the inhibitor oligonucleotide.
1. An isolated nanovesicle comprising a recombinantly expressed c-Myc inhibitor oligonucleotide, and further comprising a targeting molecule on the surface of the nanovesicle, wherein said targeting molecule is an antibody or fab-fragment thereof, and wherein the nanovesicle is produced by a producer cell recombinantly expressing L-myc.
2. An isolated nanovesicle according to claim 1 , wherein the isolated nanovesicle further comprises recombinant L-myc.
3. An isolated nanovesicle of claim 1 , wherein said oligonucleotide is an RNA molecule.
4. An isolated nanovesicle of claim 3 , wherein said oligonucleotide is an RNAi molecule.
5. An isolated nanovesicle of claim 4 , wherein said RNA is shRNA or siRNA comprising a sequence selected from the group consisting of SEQ ID NO: 93-102.
6. An isolated nanovesicle of claim 1 which is an artificial nanovesicle produced by serial extrusions of cells through microfilters and/or nanofilters.
7. An isolated nanovesicle of claim 1 and a pharmaceutically acceptable carrier.