Lipid encapsulating interfering RNA
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1. A method of introducing an interfering RNA into a cell, said method comprising contacting said cell with a nucleic acid-lipid particle comprising
(a) said interfering RNA;
(b) a cationic lipid of Formula I having the following structure:
wherein:
R 1 and R 2 are independently selected from the group consisting of: H and C 1 -C 3 alkyl; and
R 3 and R 4 are independently selected from the group consisting of alkyl groups having from about 10 to about 20 carbon atoms, wherein at least one of R 3 and R 4 comprises at least two sites of unsaturation;
(c) a non-cationic lipid; and
(d) a conjugated lipid that inhibits aggregation of particles.
2. The method of claim 1 , wherein said interfering RNA in said nucleic acid-lipid particle is resistant in aqueous solution to degradation with a nuclease.
3. The method of claim 1 , wherein said particle has a median diameter of less than about 150 nm.
4. The method of claim 1 , wherein said interfering RNA comprises a small interfering RNA (siRNA).
5. The method of claim 1 , wherein said interfering RNA is transcribed from a plasmid.
6. The method of claim 1 , wherein said non-cationic lipid is a member selected from the group consisting of dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), di stearoylphosphatidylcholine (DSPC), palmitoyloleyolphosphatidylglycerol (POPG), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, palmitoyloleoyl-phosphatidylethanolamine (POPE), 1-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), cholesterol, and a mixture thereof.
7. The method of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles is a member selected from the group consisting of a polyethyleneglycol (PEG)-lipid conjugate, a polyamide (ATTA)-lipid conjugate, and a mixture thereof.
8. The method of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles is a polyethyleneglycol (PEG)-lipid.
9. The method of claim 8 , wherein the PEG-lipid is member selected from the group consisting of a PEG-diacylglycerol, a PEG-dialkyloxypropyl, a PEG-phospholipid, a PEG-ceramide, and a mixture thereof.
10. The method of claim 1 , wherein the conjugated lipid that inhibits aggregation of particles is a polyethyleneglycol (PEG)-dialkyloxypropyl conjugate.
11. The method of claim 10 , wherein the PEG-dialkyloxypropyl conjugate is PEG-dimyristyloxypropyl (C 14 ).
12. The method of claim 1 , wherein said cell is in a mammal.
13. The method of claim 12 , wherein presence of said interfering RNA in said cell is detectable for at least 48 hours after said contacting.
14. The method of claim 12 , wherein presence of said interfering RNA in said cell is detectable for at least 24 hours after said contacting.