Synthetic dna vectors and methods of use

Abstract

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 circular DNA vector comprising a promoter operably linked to the gene, wherein the circular DNA vector lacks 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, polaxamer, 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 circular DNA vector further comprises a polyadenylation site downstream of the gene. 
     
     
         5 . The pharmaceutical composition of  claim 1 , wherein the circular DNA vector is supercoiled. 
     
     
         6 . The pharmaceutical composition of  claim 1 , wherein 70% to 99.9% of the 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 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 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.