Nonviral vectors for delivering polynucleotides to target tissues
Methods and compositions for delivering polynucleotides are provided. One embodiment provides a non-viral vector comprising a recombinant polynucleotide-binding protein comprising a protein transduction domain operably linked to a targeting signal.
1. A method for delivering a polynucleotide to one or more cells the method comprising:
contacting the one or more cells with the polynucleotide packaged with a mitochondrial transcription factor A (TFAM) polypeptide fusion protein comprising a protein transduction domain, a cell-specific targeting signal and the TFAM polypeptide wherein the TFAM polypeptide comprises two HMG box domains and binds the polynucleotide.
2. The method of claim 1 , wherein the targeting signal provides tropism for the fusion protein to a target cell type or a target cell state.
3. The method of claim 2 , wherein the targeting signal comprises a portion of bacterial Protein A sufficient to bind an Fc portion of a mammalian antibody.
4. The method of claim 3 , wherein the portion of Protein A is operably linked to an antibody specific for a cell surface antigen.
5. The method of claim 4 , wherein the antibody provides the tropism for the fusion protein to a target cell type or a target cell state.
6. The method of claim 2 , wherein the targeting signal comprises a polypeptide from a protein or a peptide comprising the targeting signal wherein the protein or the peptide is selected from the group consisting of a Surfactant protein A, a surfactant protein B, an Artery wall binding peptide, an Asialoglycoprotein, a Synthetic galactosylated ligand, a Lectin, an Anti-CD 3 antibody, an Anti-CD 5 antibody, a hyaluronic acid fragment, a Steel factor, an Anti CD 117 antibody, an EGF peptide, an Anti EGF-R antibody, a TGF-alpha, an anti ErbB2 antibody, an IgG, a basic FGF, a Folate, a Malarial circumsporozoite protein, an Anti HER2 antibody, an Insulin, an RGD peptide, a Receptor associated protein (RAP), a Synthetic ligand, a mannosylated synthetic ligand, an NGF derived synthetic peptide, an Antibody ChCE7, an Antibody OV-TL16 Fab′ fragment, an anti-PECAM antibody, an Anti-secretory component, an Anti-IgG antibody an Anti-idiotype antibody, an Anti-thrombomodulin antibody, an Anti-Tn antibody, and a Transferrin.
7. The method of claim 6 , wherein the contacting occurs in vitro or in vivo.
8. The method of claim 1 , wherein the contacting occurs in vitro or in vivo.
9. The method of claim 1 , wherein the TFAM fusion protein comprises the amino acids of SEQ ID NO: 17.
10. The method of claim 1 , wherein the TFAM fusion protein further comprises the mitochondrial localization signal of superoxide dismutase (Mn).