Nonviral vectors for delivering polynucleotides
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. Methods for modifying the genome of non-nuclear organelles are also provided.
1. A method of compensating for a Leber's Hereditary Optic Neuropathy (LHON) mutation wherein the method comprises directly contacting an eye of a subject with the LHON mutation with a mitochondrial replicon comprising an expression regulatory element operably linked to the LHON wildtype mtDNA sequence and an effective amount of a non-viral vector to package the replicon wherein the non-viral vector comprises a protein transduction domain, a mitochondrial localization signal, and mature transcription factor A, mitochondrial (TFAM).
2. A method for introducing a polynucleotide into mitochondria, the method comprising contacting one or more cells comprising the mitochondria with the polynucleotide and an effective amount of a fusion protein to package the polynucleotide,
wherein the fusion protein comprises a protein transduction domain, a mitochondrial localization signal, and a mature transcription factor A, mitochondria (TFAM),
and delivers the polynucleotide to the mitochondria.
3. The method of claim 2 wherein the polynucleotide encodes a functional protein operatively linked to an expression control sequence.
4. The method of claim 2 wherein the polynucleotide is a complete mitochondrial genome.