Mitochondrial genetic disorders are a major cause of mitochondrial diseases. It is therefore likely that mitochondrial gene therapy will be useful for the treatment of such diseases. Here, we report on the possibility of mitochondrial gene delivery in skeletal muscle using hydrodynamic limb vein (HLV) injection. The HLV injection procedure, a useful method for transgene expression in skeletal muscle, involves the rapid injection of a large volume of naked plasmid DNA (pDNA) into the distal vein of a limb. We hypothesized that the technique could be used to deliver pDNA not only to nuclei but also to mitochondria, since cytosolic pDNA that is internalized by the method may be able to overcome mitochondrial membrane. We determined if pDNA could be delivered to myofibrillar mitochondria by HLV injection by PCR analysis. Mitochondrial toxicity assays showed that the HLV injection had no influence on mitochondrial function. These findings indicate that HLV injection promises to be a useful technique for in vivo mitochondrial gene delivery.
Keywords: 4′, 6-Diamidino-2-phenylindole; CLSM; COX; Ch; DAPI; FCCP; FISH; GAPDH; Glyceraldehyde 3-phosphate dehydrogenase; HLV; Hydrodynamic limb vein injection; MELAS; MERRF; MIB; MTDR; MitoTracker Deep Red 633; Mitochondria disease; Mitochondrial gene delivery; Mitochondrial gene therapy; Mitochondrial myopathy encephalopathy, lactic acidosis and stroke-like episodes; Myoclonic epilepsy and ragged-red fiber disease; PBS (-); RLU; TMRM; Tetramethylrhodamine; carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; channel; confocal laser scanning microscopy; cytochrome c oxidase; fluorescent in situ hybridization; hydrodynamic limb vein; mitochondrial isolation buffer; pDNA; phosphate-buffered saline; plasmid DNA; q-PCR; quantitative real-time PCR; relative light units.
© 2013.