Brush polymer-assisted compaction of oligonucleotides
The disclosed subject matter relates to brush polymer-oligonucleotide conjugates comprising oligonucleotides covalently attached to the backbone of a non-cationic, sterically congested brush polymer and the use of such polymer-oligonucleotide conjugates in antisense gene regulation and as diagnostic agents.
1. A method of inhibiting expression of a gene product encoded by a target polynucleotide comprising contacting the target polynucleotide with a brush polymer-oligonucleotide conjugate comprising a plurality of oligonucleotides covalently linked to the backbone of a non-cationic, biocompatible brush polymer, wherein the oligonucleotides comprise sufficiently complementary sequences to hybridize with the target polynucleotide and inhibit expression of the target polynucleotide.
2. The method of claim 1 , wherein the target polynucleotide is a bacterial polynucleotide, viral polynucleotide or fungal polynucleotide.
3. The method of claim 1 , wherein the target polynucleotide is DNA or RNA.
4. The method of claim 1 , wherein the brush polymer-oligonucleotide conjugate comprises at least two oligonucleotides having different sequences which hybridize to different regions on the same target polynucleotide or the different oligonucleotides hybridize to different target polynucleotides which encode different gene products.
5. The method of claim 4 , wherein the brush polymer-oligonucleotide conjugate inhibits expression of more than one gene product.
6. The method of claim 1 , wherein the target polynucleotide is associated with a disease state.
7. The method of claim 6 , wherein the disease state is cancer or a bacterial, fungal or viral infection.
8. The method of claim 1 , wherein the plurality of oligonucleotides comprises an siRNA oligonucleotide which hybridizes to the target polynucleotide and initiates an RNAse activity.
9. The method of claim 1 , wherein the target polynucleotide is a double stranded polynucleotide and the plurality of oligonucleotides comprises ribozymes that hybridize to the target polynucleotide and inhibit translation of the target polynucleotide.
10. The method of claim 1 , wherein the target polynucleotide is mRNA and the brush polymer-oligonucleotide conjugate can hybridize with multiple copies of the same mRNA.
11. The method of claim 1 , wherein expression of the gene product is inhibited in vivo.
12. The method of claim 1 , wherein expression of the gene product is inhibited in vitro.
13. The method of claim 1 , wherein the brush polymer-oligonucleotide conjugate is labeled with a detectable label.
14. The method of claim 1 , wherein the brush polymer comprises polyethylene glycol (PEG) side chains.
15. The method of claim 14 , wherein the brush polymer-oligonucleotide conjugate comprises a brush polymer having PEG side chains and a plurality of single stranded or double stranded oligonucleotide molecules covalently attached to the polymer backbone.
16. The method of claim 1 , wherein the oligonucleotides are bound to the brush polymer through cleavable linkages.
17. The method of claim 1 , wherein the oligonucleotides are chemically modified or unmodified.
18. A composition comprising a plurality of oligonucleotides covalently linked to the backbone of a non-cationic, biocompatible brush polymer, and a pharmaceutically acceptable carrier, wherein the oligonucleotides comprise sufficiently complementary sequences to hybridize with a target polynucleotide and inhibit expression of the target polynucleotide.
19. The composition of claim 18 , wherein the non-cationic, biocompatible brush polymer comprises polyethylene glycol (PEG), a polysaccharide or a zwitterion polymer.
20. The composition of claim 18 , wherein the plurality of oligonucleotides is selected from the group consisting of single stranded DNA, double stranded DNA, single stranded RNA, double stranded RNA and aptamers.
21. The composition of claim 18 , wherein the oligonucleotides are chemically modified or unmodified.