Methods for arranging and packing nucleic acids for unusual resistance to nucleases and targeted delivery for gene therapy
There are disclosed compositions and methods to render nucleic acids resistant to nuclease digestion while maintaining sequence selective hybridization competency. The approach relies on utilizing nucleic acids as the polar head group of a nucleic acid-polymer amphiphile in order to assemble well-defined, discrete micellar nanoparticles. Dense packing of nucleic acid in the micelle corona allows for hybridization of complementary oligonucleotides while prohibiting enzymatic degradation.
1. A micelle comprising a plurality of nucleic acid conjugates, each of said nucleic acid conjugates comprising:
a first oligonucleotide comprising up to about 200 bases, wherein said first oligonucleotide is complementary to a target sequence associated with aberrant physiological activity, and
a cyclic olefin polymer covalently bound to said first oligonucleotide through a first linker, wherein said first linker comprises an alkylene of at least 12 carbons;
wherein each of said conjugates forms nanoparticulate micelles having a hydrophobic core and a hydrophilic shell;
wherein said hydrophobic core comprises a plurality of cyclic olefin polymer moieties; and
wherein said hydrophilic shell comprises a high density of said first oligonucleotide.
2. The micelle of claim 1 , wherein said first oligonucleotide is a DNA, an RNA or a locked nucleic acid (LNA).
3. The micelle of claim 1 , wherein said cyclic olefin polymer is covalently linked, directly or indirectly, to said first oligonucleotide via an amide bond.
4. The micelle of claim 1 , wherein said high density of said first oligonucleotide comprises at least 20 oligonucleotides per micellar nanoparticle.
5. The micelle of claim 1 , further comprising a second oligonucleotide which is at least partially complementary to said first oligonucleotide.
6. A formulation comprising the micelle of claim 1 in a pharmaceutically acceptable carrier therefore.
7. A method for delivery of a micelle into a target cell, said method comprising administering an effective amount of a formulation according to claim 6 to a subject in need thereof.
8. A method to impart endonuclease resistance to a micelle comprising a plurality of nucleic acid conjugates of claim 1 , said method comprising incorporating said nucleic acid conjugates into a micelle according to claim 1 prior to exposure of said micelle to an endonuclease, thereby imparting endonuclease resistance to said micelle comprising a plurality of nucleic acid conjugates.