Mini-dystrophin nucleic acid sequences
The present invention relates to compositions and methods for expressing mini-dystrophin peptides. In particular, the present invention provides compositions comprising nucleic acid sequences that are shorter than wild-type dystrophin cDNA and that express mini-dystrophin peptides that function in a similar manner as wild-type dystrophin proteins. The present invention also provides compositions comprising mini-dystrophin peptides, and methods for expressing mini-dystrophin peptides in target cells.
1. A composition comprising nucleic acid encoding a mini-dystrophin peptide, wherein said mini-dystrophin peptide comprises a spectrin-like repeat domain comprising 4 dystrophin spectrin-like repeats, wherein said mini-dystrophin peptide contains no more than 4 dystrophin spectrin-like repeats.
2. The composition of claim 1 , wherein said dystrophin spectrin-like repeats are human dystrophin spectrin-like repeats.
3. The composition of claim 1 , wherein said mini-dystrophin-peptide is capable of increasing a measurable muscle value in a DMD animal model by at least 20% of the wild type value, wherein said measurable muscle value is a diaphragm specific force value.
4. The composition of claim 3 , wherein said mini-dystrophin peptide is capable of increasing said diaphragm specific force value in a DMD animal model by at least 30% of the wild-type value.
5. The composition of claim 1 , wherein said nucleic acid comprises an expression vector.
6. The composition of claim 1 , wherein said nucleic acid comprises spectrin-like repeat encoding sequences.
7. The composition of claim 6 , wherein said spectrin-like repeat encoding sequences are precise spectrin-like repeat encoding sequences.
8. The composition of claim 1 , wherein said nucleic acid comprises an actin-binding domain encoding sequence.
9. The composition of claim 8 , wherein said actin binding domain comprises at least a portion of SEQ ID NO:6.
10. The composition of claim 1 , wherein said nucleic acid comprises a β-dystroglycan binding domain.
11. The composition of claim 10 , wherein said β-dystroglycan binding domain comprises at least a portion of a dystrophin hinge 4 encoding sequence, and at least a portion of a dystrophin cysteine-rich domain encoding sequence.
12. The composition of claim 6 , wherein said spectrin-like repeat encoding sequences are selected from the group consisting of SEQ ID NOS:8-10, 12-27, and 29-33.
13. The composition of claim 1 , wherein said nucleic acid contains less than 75% of a wild type dystrophin 5′ untranslated region.
14. The composition of claim 1 , wherein said mini-dystrophin peptide further comprises a substantially deleted dystrophin C-terminal domain.
15. The composition of claim 1 , wherein said nucleic acid contains less than 50% of a dystrophin 3′ untranslated region.
16. A composition comprising nucleic acid encoding a mini-dystrophin peptide, wherein said mini-dystrophin peptide comprises i) a spectrin-like repeat domain comprising 4 dystrophin spectrin-like repeats, ii) an actin-binding domain, and iii) a β-dystroglycan binding domain; and wherein said mini-dystrophin peptide contains no more than 4 dystrophin spectrin-like repeats.
17. The composition of claim 16 , wherein said mini-dystrophin-peptide is capable of altering increasing a measurable muscle value in a DIVID animal model by at least 20% of the wild type value wherein said measurable muscle value is a diaphragm specific force value.
18. The composition of claim 17 , wherein said mini-dystrophin peptide is capable of increasing said diaphragm specific force value in a DMD animal model by at least 30% of the wild-type value.
19. The composition of claim 16 , wherein said nucleic acid is less than 5.0 kb in length.
20. A composition comprising nucleic acid encoding a mim-dystrophin peptide, wherein said mini-dystrophin peptide comprises a spectrin-like repeat domain comprising 8 dystrophin spectrin-like repeats, wherein said mini-dystrophin peptide contains no more than 8 dystrophin spectrin-like repeats.
21. The composition of claim 20 , wherein said dystrophin spectrin-like repeats are human dystrophin spectrin-like repeats.
22. The composition of claim 20 , wherein said mini-dystrophin-peptide is capable of altering a measurable muscle value in a DMD animal model by at least 20% of the wild type value.
23. The composition of claim 16 , wherein said nucleic acid contains less than 50% of a dystrophin 3+ untranslated region.
24. The composition of claim 1 , wherein said mini-dystrophin peptide further comprises dystrophin hinge region 1 and dystrophin hinge region 4.
25. The composition of claim 24 , wherein said mini-dystrophin further comprises dystrophin hinge region 2 or dystrophin hinge region 3.
26. The composition of claim 16 , wherein said mini-dystrophin peptide further comprises dystrophin hinge region 1 and dystrophin hinge region 4.
27. The composition of claim 26 , wherein said mini-dystrophin further comprises dystrophin hinge region 2 or dystrophin hinge region 3.
28. The composition of claim 1 , wherein said nucleic acid is less than 5.0 kb in length.
29. The composition of claim 5 , wherein said expression vector comprises an adeno-associated viral sequence, and wherein said nucleic acid comprises a promoter.
30. The composition of claim 29 , wherein said promoter is an MCK promoter.
31. The composition of claim 16 , wherein said nucleic acid comprises an adeno-associated viral sequence and a promoter.
32. The composition of claim 31 , wherein said promoter comprises an MCK promoter.
33. The composition of claim 1 , wherein said 4 dystrophin spectrin-like repeats are selected from the group consisting of: dystrophin spectrin-like repeat number 1, dystrophin spectrin-like repeat number 2, dystrophin spectrin-like repeat number 3, dystrophin spectrin-like repeat number 22, dystrophin spectrin-like repeat number 23, and dystrophin spectrin like repeat number 24.
34. The composition of claim 16 , wherein said 4 dystrophin spectrin-like repeats are selected from the group consisting of: dystrophin spectrin-like repeat number 1, dystrophin spectrin-like repeat number 2, dystrophin spectrin-like repeat number 3, dystrophin spectrin-like repeat number 22, dystrophin speetrin-like repeat number 23, and dystrophin spectrin like repeat number 24.