Reducing cellular dysfunction caused by mitochondrial gene mutations
View Patent ↗A synthetic, nuclear-encoded ND4 gene was linked to a mitochondrial targeting sequence and a FLAG epitope tag. This fusion construct was inserted into a rAAV vector. The ND4 fusion protein was expressed and imported into the mitochondria of cells harboring a mitochondrial DNA mutation (G11778A), where it restored cellular respiration.
1. An isolated nucleic acid comprising a nucleotide sequence encoding a functional ND4 mitochondrial protein, wherein the nucleotide sequence comprises the sequence of SEQ ID NO:1.
2. The isolated nucleic acid of claim 1 , wherein the nucleic acid is comprised within an expression vector.
3. The isolated nucleic acid of claim 2 , wherein the expression vector is a plasmid.
4. The isolated nucleic acid of claim 1 , wherein the nucleic acid is comprised within an rAAV virion.
5. The isolated nucleic acid of claim 1 , wherein the nucleic acid further comprises a nucleotide sequence encoding a mitochondrial targeting sequence.
6. The isolated nucleic acid of claim 1 , wherein the nucleic acid further comprises a promoter operably linked to the nucleotide sequence.
7. The isolated nucleic acid of claim 1 , wherein the nucleic acid further comprises an enhancer element.
8. The isolated nucleic acid of claim 1 , wherein the nucleic acid further comprises a polyA tail.
9. An isolated cell into which has been introduced a nucleic acid comprising a nucleotide sequence encoding a functional ND4 mitochondrial protein wherein the nucleotide sequence comprises the sequence of SEQ ID NO: 1.
10. The cell of claim 9 , wherein the cell is a human cell.
11. The cell of claim 10 , wherein the cell is a human nerve cell.