Method of delivering RNA interference and uses thereof
The invention provides a method of RNA interference, which comprises contacting the cell with a fusion protein-double stranded RNA complex, the complex comprising the double stranded RNA segment containing a double stranded RNA of interest and a fusion protein, the fusion protein comprising (1) a targeting moiety, which will specifically binds to a site on a target cell, and (2) a binding moiety, which will bind to the double stranded RNA, wherein the double stranded RNA segment initiates RNA interference in the cell.
1. A composition that selectively binds to an antigen on the surface of a target cell and selectively initiates RNA interference in the cell comprising:
a) an siRNA molecule comprising a double stranded RNA segment of about 15-50 base pairs in length, wherein one of one strands is complementary and the other strand identical to a RNA interference target RNA; and
b) a fusion protein, comprising (1) an antibody that binds specifically to an antigen on the surface of a target cell, and (2) a protamine protein, or fragment thereof, capable of binding to double stranded RNA without chemical conjugation.
2. The composition of claim 1 , wherein the antibody is an antibody to a viral envelope protein, a cellular receptor, or an extracellular domain of an activated receptor.
3. The composition of claim 1 , wherein the antibody is a single chain antibody, a Fab portion of an antibody or a (Fab') 2 segment.
4. The composition of claim 3 , wherein the antibody recognizes Gp120 or gp160.
5. The composition of claim 1 , wherein the-protamine protein, or portion thereof, is fused to the carboxyl portion of the antibody.
6. The composition of claim 1 , wherein the siRNA targets mRNA encoding c-myc, VEGF, CD4, CCR5, gag, MDM2, Apex, Ku70, or ErbB2.
7. The composition of claim 1 , wherein the cell is selected from the group consisting of hepatocytes, myocytes, neural cells, lipocytes, lymphocytes, macrophages, cardiac cells, endothelial cells, and epithelial cells.
8. The composition of claim 1 , wherein the target cell is a malignant cell.
9. The composition of claim 8 , wherein the malignant cell is selected from the group consisting of a lung cancer cell, a retinal cancer cell, a breast cancer cell, a ovarian cancer cell, a prostate cancer cell, a head and neck cancer cell, a lymphoma cell, a melanoma cell, a glioma cell, a bladder cancer cell, a genital-urinary cancer cell, a stomach cancer cell, a pancreatic cancer cell, a liver cancer cell, a kidney cancer cell, and a gastrointestinal cancer cell.
10. The composition of claim 1 , comprising further comprising a pharmaceutical carrier.
11. The composition of claim 10 , wherein the pharmaceutical carrier is formulated for oral, rectal, vaginal, parenteral, topical, or intravenous administration.
12. The composition of claim 1 , wherein the protamine fragment thereof is encoded by a nucleic acid having a sequence selected from the group consisting of:
SEQ ID NO:1, SEQ ID NO:2; SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, and nucleic acids 10-76 of SEQ ID NO:1.
13. A kit comprising the composition of claim 1 .
14. A kit comprising:
a) an siRNA molecule comprising double stranded RNA segments of about 15-50 base pairs in length; and
b) a fusion protein, comprising (1) an antibody that binds specifically to an antigen on the surface of a target cell, and (2) a protamine protein, or fragment thereof, capable of binding to double stranded RNA without chemical conjugation.
15. The kit of claim 14 , further comprising a plurality of different siRNA molecules.