Interfering RNA delivery system and uses thereof
The invention provides a delivery system comprising a cell penetrating peptide, 10 histidines, and an interfering RNA molecule. The system can be used for delivering interfering RNA molecules into a cell in vivo or in vitro. Therapeutic uses for the delivery system are also provided.
1. A composition comprising an interfering RNA molecule covalently linked to a YM3-10H peptide, wherein the YM3-10H peptide comprises:
(SEQ ID NO: 2)
CHHHHHTHRLPRRRRRRHHHHHC;
(SEQ ID NO: 3)
HHHHHTHRLPRRRRRRHHHHHC;
(SEQ ID NO: 5)
GGTHRLPRRRRRRHHHHHHHHHHC;
(SEQ ID NO: 6)
THRLPRRRRRRGGHHHHHHHHHHC;
(SEQ ID NO: 7)
THRLPRRRRRRHHHHHGGHHHHHC;
or
(SEQ ID NO: 9)
CTHRLPRRRRRRHHHHHHHHHHC.
2. The composition of claim 1 , wherein the YM3-10H peptide has an amino-terminal cysteine residue.
3. The composition of claim 1 , wherein the YM3-10H peptide has a carboxy-terminal cysteine residue.
4. The composition of claim 1 , wherein the YM3-10H peptide has an amino-terminal cysteine residue and a carboxy-terminal cysteine residue.
5. The composition of claim 1 , wherein the composition further comprises a second YM3-10H peptide wherein the second YM3-10H peptide comprise the amino acid sequence of SEQ ID NO: 1 and at least ten histidine residues, and wherein the first YM3-10H peptide and the second YM3-10H peptide form a concatemer.
6. The composition of claim 5 , wherein the concatemer comprises the amino acid sequence of SEQ ID NO: 10.
7. The composition of claim 1 , wherein the YM3-10H peptide consists of:
(SEQ ID NO: 2)
CHHHHHTHRLPRRRRRRHHHHHC;
(SEQ ID NO: 3)
HHHHHTHRLPRRRRRRHHHHHC;
(SEQ ID NO: 5)
GGTHRLPRRRRRRHHHHHHHHHHC;
(SEQ ID NO: 6)
THRLPRRRRRRGGHHHHHHHHHHC;
(SEQ ID NO: 7)
THRLPRRRRRRHHHHHGGHHHHHC;
or
(SEQ ID NO: 9)
CTHRLPRRRRRRHHHHHHHHHHC.
8. The composition of claim 1 , wherein the interfering RNA molecule is covalently linked to the YM3-10H peptide via a spacer.
9. The composition of claim 1 , wherein the interfering RNA molecule is a short interfering RNA (SiRNA), a microRNA (miRNA), or a short hairpin RNA (shRNA).
10. The composition of claim 1 , further comprising a pharmaceutically acceptable carrier.
11. The composition of claim 1 , wherein the YM3-10H comprises CHHHHHTHRLPRRRRRRHHHHHC (SEQ ID NO: 2).
12. The composition of claim 1 , wherein the YM3-10H comprises HHHHHTHRLPRRRRRRHHHHHC (SEQ ID NO: 3).
13. The composition of claim 1 , wherein the YM3-10H comprises GGTHRLPRRRRRRHHHHHHHHHHC (SEQ ID NO: 5).
14. The composition of claim 1 , wherein the YM3-10H comprises THRLPRRRRRRGGHHHHHHHHHHC (SEQ ID NO: 6).
15. The composition of claim 1 , wherein the YM3-10H comprises THRLPRRRRRRHHHHHGGHHHHHC (SEQ ID NO: 7).
16. The composition of claim 1 , wherein the YM3-10H comprises CTHRLPRRRRRRHHHHHHHHHHC (SEQ ID NO: 9).
17. A method of delivering an interfering RNA molecule into a cell, the method comprising administering the composition of claim 1 to a cell.
18. The method of claim 17 , wherein the interfering RNA molecule is a siRNA, a miRNA, or a shRNA.
19. The method of claim 17 , wherein the YM3-10H peptide consists of:
(SEQ ID NO: 2)
CHHHHHTHRLPRRRRRRHHHHHC;
(SEQ ID NO: 3)
HHHHHTHRLPRRRRRRHHHHHC;
(SEQ ID NO: 5)
GGTHRLPRRRRRRHHHHHHHHHHC;
(SEQ ID NO: 6)
THRLPRRRRRRGGHHHHHHHHHHC;
(SEQ ID NO: 7)
THRLPRRRRRRHHHHHGGHHHHHC;
or
(SEQ ID NO: 9)
CTHRLPRRRRRRHHHHHHHHHHC.