Synthetic DNA vectors and methods of use
Provided herein are isolated DNA vectors comprising a heterologous gene, wherein the DNA vector is devoid of bacterial plasmid DNA and/or bacterial signatures, which can abrogate persistence in vivo. The invention also features pharmaceutical compositions (non-immunogenic pharmaceutical compositions) including the DNA vectors of the invention, which can be used for induction of long-term, episomal expression of a heterologous gene in a subject. The invention involves methods of treating a subject by administering the DNA vectors of the invention, including methods of treating disorders associated with a defect in a target gene.
1. A pharmaceutical composition for delivery of a gene in a subject, wherein the pharmaceutical composition comprises a synthetic circular DNA vector comprising a promoter operably linked to the gene, wherein the synthetic circular DNA vector lacks an origin of replication, a drug resistance gene, and a site-specific recombination recognition site, wherein the promoter is capable of inducing expression of the gene in the subject.
2. The pharmaceutical composition of claim 1 , further comprising a delivery vehicle selected from liposomes, nanoparticles, microparticles, microspheres, lipid particles, vesicles, polyaxamer, and polycationic material, and combinations thereof.
3. The pharmaceutical composition of claim 1 , wherein the promoter is substantially devoid of CpG islands.
4. The pharmaceutical composition of claim 1 , wherein the synthetic circular DNA vector further comprises a polyadenylation site downstream of the gene.
5. The pharmaceutical composition of claim 1 , wherein the synthetic circular DNA vector is supercoiled.
6. The pharmaceutical composition of claim 1 , wherein 70% to 99.9% of the synthetic circular DNA vector in the pharmaceutical composition is monomeric.
7. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is formulated to be administered intravenously, intramuscularly, intradermally, intracerebrally, intrathecally, intratumorally, intratracheally, intrapleurally, or by inhalation.
8. The pharmaceutical composition of claim 1 , wherein the gene is at least 5 Kb in length.
9. The pharmaceutical composition of claim 1 , wherein the synthetic circular DNA vector is substantially devoid of CpG islands.
10. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition does not cause significant increases of cytokine levels when administered to a subject.
11. The pharmaceutical composition of claim 1 , wherein the gene encodes a polypeptide.
12. The pharmaceutical composition of claim 11 , wherein the polypeptide is a replacement polypeptide.
13. The pharmaceutical composition of claim 1 , wherein the gene encodes a therapeutic nucleic acid.
14. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition comprises a unit dose of the synthetic circular DNA vector of from 10 μg to 10 mg.
15. The pharmaceutical composition of claim 1 , wherein the pharmaceutical composition is substantially devoid of an immunogenic component.
16. The pharmaceutical composition of claim 15 , wherein the immunogenic component comprises endotoxin, bacterial contaminants, flagellin, lipoteichoic acid, peptidoglycan, or a combination thereof.
17. The pharmaceutical composition of claim 1 , wherein the gene is a human gene.