Genetic correction of mutated genes
Disclosed herein are transcription activator-like effector nuclease (TALEN)-related compositions and methods of using said TALENs for correcting mutant genes.
1. An isolated polynucleotide comprising a nucleotide sequence encoding an engineered transcription activator-like effector nuclease (TALEN) protein that binds to a dystrophin gene, wherein the TALEN protein binds upstream from a premature stop codon on the dystrophin gene, downstream from a premature stop codon on the dystrophin gene, a region in exon 51 of the dystrophin gene, or a region in the 5′ UTR of the dystrophin gene, and wherein the TALEN protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, and SEQ ID NO: 15.
2. The isolated polynucleotide of claim 1 , wherein the TALEN protein binds to a nucleotide sequence comprising one of SEQ ID NOs: 16-46, 50, 52, 58, and 59, or a complement thereof.
3. The isolated polynucleotide of claim 1 , wherein the TALEN protein comprises a nuclease.
4. The isolated polynucleotide of claim 3 , wherein the nuclease comprises FokI.
5. The isolated polynucleotide of claim 1 , wherein the TALEN protein comprises 15-19 repeat variable diresidue (RVD) modules.
6. A vector comprising the isolated polynucleotide of claim 1 .
7. The vector of claim 6 , wherein the TALEN protein binds to a nucleotide sequence comprising one of SEQ ID NOs: 16-46, 50, 52, 58, and 59, or a complement thereof.
8. The vector of claim 6 , wherein the TALEN protein comprises a nuclease.
9. The vector of claim 8 , wherein the nuclease comprises FokI.
10. The vector of claim 6 , wherein the TALEN protein comprises 15-19 repeat variable diresidue (RVD) modules.
11. A cell comprising the isolated polynucleotide of claim 1 .
12. The cell of claim 11 , wherein the TALEN protein binds to a nucleotide sequence comprising one of SEQ ID NOs: 16-46, 50, 52, 58, and 59, or a complement thereof.
13. The cell of claim 11 , wherein the TALEN protein comprises a nuclease.
14. The cell of claim 13 , wherein the nuclease comprises FokI.
15. The cell of claim 11 , wherein the TALEN protein comprises 15-19 repeat variable diresidue (RVD) modules.