Compositions and methods for the treatment of disorders related to frataxin deficiency

Abstract

The disclosure relates to compositions and methods for, inter alia, altering, e.g., enhancing, the level of frataxin protein via delivery using an adeno-associated viral (AAV) capsid variant. The compositions and methods of the present disclosure are useful, inter alia, in the treatment of subjects who have, have been diagnosed with, or suspected of having a disorder associated with frataxin (FXN) deficiency, e.g., Friedreich's Ataxia.

Description

CN Title: Compositions and Methods for Treating Disorders Associated with Ataxin Deficiency CN Abstract: This disclosure relates to compositions and methods, particularly for altering, for example enhancing, ataxin levels via delivery using an adeno-associated virus (AAV) capsid variant. The compositions and methods disclosed herein are particularly useful for treating subjects who have, have been diagnosed with, or are suspected of having disorders associated with ataxin (FXN) deficiency, such as Friedreich ataxia. 1 Abstract

Claims

CLAIMS What is claimed is:

1. An adeno-associated virus (AAV) particle comprising: a) an AAV capsid variant comprising an amino acid sequence having the following formula: [N1]-[N2]-[N3], wherein: (i) optionally [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G; (ii) [N2] comprises the amino acid sequence of SPH; and (iii) [N3] comprises X4, X5, and X6, wherein at least one of X4, X5, or X6 is a basic amino acid; and b) a viral genome comprising a frataxin (FXN)-encoding sequence.

2. The AAV particle of claim 1, wherein the amino acid sequence [N1]-[N2]-[N3] is in hypervariable loop IV of the AAV capsid variant.

3. The AAV particle of claim 1 or claim 2, wherein the AAV capsid variant is an AAV9 capsid variant.

4. The AAV particle of any one of claims 1-3, wherein [N1] comprises X1, X2, and X3, wherein at least one of X1, X2, or X3 is G.

5. The AAV particle of any one of claims 1-4, wherein [N2]-[N3] comprises the amino acid sequence of SPHSKA (SEQ ID NO: 941).

6. An adeno-associated virus (AAV) particle comprising a viral genome comprising a frataxin (FXN)- encoding sequence and an AAV9 capsid variant comprising the amino acid sequence of SPHSKA (SEQ ID NO: 941).

7. The AAV particle of claim 6, wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is in hypervariable loop IV of the AAV9 capsid variant.

8. The AAV particle of claim 6 or claim 7, wherein the amino acid sequence of SPHSKA (SEQ ID NO: 941) is present immediately subsequent to an amino acid position corresponding to position 455 of SEQ ID NO: 4 or SEQ ID NO:

36.

9. The AAV particle of any one of claims 6-8, wherein the AAV9 capsid variant further comprises one, two, or all of: an N at an amino acid position corresponding to position 452, an E at an aminoacid position corresponding to position 451, and / or a V at an amino acid position corresponding to position 453 of SEQ ID NO:

4.

10. The AAV particle of claim any one of claims 6-9, wherein the AAV9 capsid variant comprises the amino acid sequence of KTENVSGSPHSKAQNQQT (SEQ ID NO: 3272).

11. The AAV particle of any one of claims 6-10, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence having at least 90% identity to positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 90% identity to positions 203-742 of SEQ ID NO:

4.

12. The AAV particle of any one of claims 6-11, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 95% identity to SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence having at least 95% identity to positions 138-742 SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 95% identity to positions 203-742 of SEQ ID NO:

4.

13. The AAV particle of any one of claims 6-12, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 99% identity to SEQ ID NO: 4; (ii) a VP2 protein comprising an amino acid sequence having at least 99% identity to positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 99% identity to positions 203-742 of SEQ ID NO:

4.

14. The AAV particle of any one of claims 6-13, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 4.

15. The AAV particle of any one of claims 6-13, wherein the AAV9 capsid variant comprises: (i) the amino acid sequence of SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to an amino acid position corresponding to position 455 of SEQ ID NO: 4; (ii) an E at an amino acid position corresponding to position 451 and a V at an amino acid position corresponding to position 453 of SEQ ID NO: 4; and (iii) no other modifications relative to wild type AAV9.

16. The AAV particle of any one of claims 6-8, wherein the AAV9 capsid variant further comprises one, two, or all of: an E at an amino acid position corresponding to position 451, an R at an amino acid position corresponding to position 452, and / or a V at an amino acid position corresponding to position 453 of SEQ ID NO:

36.

17. The AAV particle of any one of claims 6-8 and 16, wherein the AAV9 capsid variant comprises the amino acid sequence of KTERVSGSPHSKAQNQQT (SEQ ID NO: 3589).

18. The AAV particle of any one of claims 6-8, 16, and 17, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence having at least 90% identity to positions 138-742 SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 90% identity to positions 203-742 of SEQ ID NO:

36.

19. The AAV particle of any one of claims 6-8 and 16-18, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising an amino acid sequence having at least 95% identity to SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence having at least 95% identity to positions 138-742 SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 95% identity to positions 203-742 of SEQ ID NO:

36.

20. The AAV particle of any one of claims 6-8 and 16-19, wherein the AAV9 capsid variant comprises:(i) a VP1 protein comprising an amino acid sequence having at least 99% identity to SEQ ID NO: 36; (ii) a VP2 protein comprising an amino acid sequence having at least 99% identity to positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising an amino acid sequence having at least 99% identity to positions 203-742 of SEQ ID NO:

36.

21. The AAV particle of any one of claims 6-8 and 16-20, wherein the AAV9 capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:

36.

22. The AAV particle of any one of claims 6-8 and 16-20, wherein the AAV9 capsid variant comprises: (i) the amino acid sequence SPHSKA (SEQ ID NO: 941), wherein the amino acid sequence is present immediately subsequent to an amino acid position corresponding to position 455 of SEQ ID NO: 36; (ii) an E at an amino acid position corresponding to position 451, an R at an amino acid position corresponding to position 452, and a V at an amino acid position corresponding to position 453 of SEQ ID NO: 36; and (iii) no other modifications relative to wild type AAV9.

23. The AAV particle of any one of claims 1-4, wherein [N1]-[N2]-[N3] is present immediately subsequent to a position corresponding to the amino acid position 452 of SEQ ID NO: 982; and wherein the AAV capsid variant comprises an amino acid sequence at least 90% identical, e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical, to the amino acid sequence of SEQ ID NO: 982, e.g., to positions 203-742 of SEQ ID NO:

982.

24. The AAV particle of claim 23, wherein [N1] comprises GHD.

25. The AAV particle of claim 23 or claim 24, wherein [N1] comprises the amino acid G at a position corresponding to position 453, the amino acid H at position 454, and the amino acid D at position 455 of SEQ ID NO: 138 or SEQ ID NO: 982.

26. The AAV particle of any one of claims 23-25, wherein [N3] comprises KSG.

27. The AAV particle of any one of claims 23-26, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity to SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity to positions 138-742 SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity to positions 203-742 of SEQ ID NO:

982.

28. The AAV particle of any one of claims 23-27, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity to SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity to positions 138-742 SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 95% identity to positions 203-742 of SEQ ID NO:

982.

29. The AAV particle of any one of claims 23-28, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity to SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity to positions 138-742 SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 99% identity to positions 203-742 of SEQ ID NO:

982.

30. The AAV particle of any one of claims 23-29, wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:

982.

31. The AAV particle of any one of claims 1-30, wherein the FXN protein encoded by the FXN- encoding sequence is not a cynomolgus FXN protein.

32. The AAV particle of any one of claims 1-31, wherein the FXN-encoding sequence encodes a human FXN protein.

33. The AAV particle of any one of claims 1-32, wherein the FXN-encoding sequence comprises SEQ ID NO: 1824 or a nucleotide sequence that is at least 80% identical (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1824.

34. The AAV particle of any one of claims 1-33, wherein the FXN-encoding sequence comprises SEQ ID NO: 1824.

35. The AAV particle of any one of claims 1-34, wherein the viral genome further comprises a promoter operably linked to the FXN-encoding sequence.

36. The AAV particle of claim 35, wherein the promoter comprises a human elongation factor 1α- subunit (EF1α) promoter, a cytomegalovirus (CMV) immediate-early enhancer and / or promoter, a chicken β-actin (CBA) promoter, a CAG promoter, a β glucuronidase (GUSB) promoter, a ubiquitin C (UBC) promoter, a neuron-specific enolase (NSE) promoter, a platelet-derived growth factor (PDGF) promoter, a platelet-derived growth factor B-chain (PDGF-β) promoter, a intercellular adhesion molecule 2 (ICAM-2) promoter, a synapsin (Syn) promoter, a methyl-CpG binding protein 2 (MeCP2) promoter, a Ca2+ / calmodulin-dependent protein kinase II (CaMKII) promoter, a metabotropic glutamate receptor 2 (mGluR2) promoter, a neurofilament light chain (NFL) or neurofilament heavy chain (NFH) promoter, a β-globin minigene nβ2 promoter, a preproenkephalin (PPE) promoter, a enkephalin (Enk) and excitatory amino acid transporter 2 (EAAT2) promoter, a glial fibrillary acidic protein (GFAP) promoter, a myelin basic protein (MBP) promoter, a cardiovascular promoter (e.g., αMHC, cTnT, and CMV-MLC2k), a liver promoter (e.g., hAAT, TBG), a skeletal muscle promoter (e.g., desmin, MCK, C512), or a functional fragment or truncation of any of the foregoing.

37. The AAV particle of claim 35 or claim 36, wherein the promoter is a CMV promoter or CBA promoter, or a functional fragment or truncation thereof.

38. The AAV particle of any one of claims 35-37, wherein the promoter is a truncated CBA promoter.

39. The AAV particle of claim 38, wherein the truncated CBA promoter is 50-400 nucleotides in length, e.g., 100-332 nucleotides in length.

40. The AAV particle of any one of claims 35-39, wherein the promoter comprises or consists of the nucleotide sequence of any one of SEQ ID NOs: 1738, 1740, and 1742 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to any one of SEQ ID NOs: 1738, 1740, and 1742.

41. The AAV particle of claim 37, wherein the promoter is a truncated CMV promoter.

42. The AAV particle of claim 41, wherein the truncated CMV promoter is 109 nucleotides in length.

43. The AAV particle of claim 41 or claim 42, wherein the promoter comprises or consists of the nucleotide sequence of SEQ ID NO: 1750 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1750.

44. The AAV particle of any one of claims 1-43, wherein the viral genome further comprises a miRNA (miR) binding site that modulates expression of the encoded FXN protein in a cell or tissue of the liver.

45. The AAV particle of claim 44, wherein the viral genome comprises 3 copies of the miR binding site.

46. The AAV particle of claim 45, wherein the 3 copies of the miR binding site are identical.

47. The AAV particle of claim 45 or claim 46, wherein the 3 copies of the miR binding site are continuous.

48. The AAV particle of any one of claims 44-47, wherein the miR binding site is a miR122 binding site.

49. The AAV particle of any one of claims 44-48, wherein: the miR122 binding site comprises the nucleotide sequence of SEQ ID NO: 1827 or a sequence having one, two, three, or at most four substitutions relative to SEQ ID NO: 1827; or the 3 copies of continuous miR122 binding sites (miR122 binding site series) comprises the nucleotide sequence of SEQ ID NO: 1826 or a sequence having one, two, three, four, five, six, seven, eight, nine, or at most ten substitutions relative to SEQ ID NO: 1826.

50. The AAV particle of any one of claims 1-49, wherein the viral genome further comprises at least one inverted terminal repeat (ITR) region.

51. The AAV particle of claim 50, wherein the at least one ITR region comprises an AAV2 ITR.

52. The AAV particle of claim 50 or claim 51, wherein the viral genome comprises a 5’ ITR region and a 3’ ITR region.

53. The AAV particle of claim 52, wherein the 5’ ITR region and the 3’ ITR region is each an AAV2 ITR.

54. The AAV particle of claim 53, wherein: the 5' ITR region comprises the nucleotide sequence of SEQ ID NO: 1811 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and / or the 3' ITR region comprises the nucleotide sequence of SEQ ID NO: 1812 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

55. The AAV particle of any one of claims 1-54, wherein the viral genome further comprises an intron / exon region comprising an intron region and / or an exon region, wherein the intron / exon region comprises: an immediate-early 1 (ie1) intron region and / or a human beta-globin (hBglobin) intron region; and / or an ie1 exon region and / or an hBglobin exon region.

56. The AAV particle of claim 55, wherein the intron region comprises: an ie1 intron 1 comprising of the nucleotide sequence of SEQ ID NO: 1819 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and / or a hBglobin intron 2 comprising the nucleotide sequence of SEQ ID NO: 1820 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

57. The AAV particle of claim 55 or claim 56, wherein the exon region comprises:an ie1 exon region comprising the nucleotide sequence of SEQ ID NO: 1817 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and / or an hBglobin exon region comprising the nucleotide sequence of SEQ ID NO: 1821 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

58. The AAV particle of any one of claims 1-57, wherein the viral genome further comprises a polyadenylation (polyA) region.

59. The AAV particle of claim 58, wherein the polyA region comprises a human growth hormone (hGH) polyA region, optionally wherein the polyA region comprises the nucleotide sequence of SEQ ID NO: 1828 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

60. The AAV particle of any one of claims 1-32, wherein the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR) region; (ii) a promoter; (iii) the FXN-encoding sequence, wherein the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a nucleotide sequence that is at least 80% identical (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1824; and (iv) a 3’ ITR region.

61. The AAV particle of any one of claims 1-32, wherein the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR) region; (ii) a promoter; (iii) an intron and / or exon region; (iv) the FXN-encoding sequence, wherein the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a nucleotide sequence that is at least 80% identical (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1824; (v) at least one miR122 binding site; and (vi) a 3’ ITR region.

62. The AAV particle of any one of claims 1-32, wherein the viral genome comprises:(i) a 5’ inverted terminal repeat (ITR) region; (ii) a promoter; (iii) an intron and / or exon region; (iv) the FXN-encoding sequence, wherein the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a nucleotide sequence that is at least 80% identical (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1824; (v) at least one miR122 binding site; (vi) a polyadenylation (polyA) region; and (vii) a 3’ ITR region.

63. The AAV particle of any one of claims 1-32, wherein the viral genome comprises: (i) a 5’ inverted terminal repeat (ITR) region; (ii) a promoter; (iii) an intron and / or exon region; (iv) the FXN-encoding sequence, wherein the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a nucleotide sequence that is at least 80% identical (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to SEQ ID NO: 1824; (v) at least one miR122 binding site; (vi) a polyadenylation (polyA) region; (vii) a filler sequence; and (viii) a 3’ ITR region.

64. The AAV particle of claim 62 or claim 63, wherein: (i) the 5’ ITR region comprises the nucleotide sequence of SEQ ID NO: 1811 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (ii) the promoter consists of the nucleotide sequence of SEQ ID NO: 1742 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iii) the intron / exon region comprises the nucleotide sequence of SEQ ID NO: 1816 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto;(v) the at least one miR122 binding site comprises a miR122 binding site series comprising SEQ ID NO: 1826 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) the polyA region comprises the nucleotide sequence of SEQ ID NO: 1828 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and / or (vii) the 3’ ITR region comprises SEQ ID NO: 1812 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

65. The AAV particle of claim 64, wherein the viral genome further comprises a filler sequence comprising the nucleotide sequence of SEQ ID NO: 1841 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto, optionally wherein the filler sequence is positioned 3’ to the polyA region and 5’ to the 3’ ITR.

66. The AAV particle of claim 62 or claim 63, wherein: (i) the 5’ ITR region comprises the nucleotide sequence of SEQ ID NO: 1811 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (ii) the promoter consists of the nucleotide sequence of SEQ ID NO: 1750 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iii) the intron / exon region comprises the nucleotide sequence of SEQ ID NO: 1816 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (v) the at least one miR122 binding site comprises a miR122 binding site series comprising SEQ ID NO: 1826 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) the polyA region comprises the nucleotide sequence of SEQ ID NO: 1828 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and / or(vii) the 3’ ITR region comprises SEQ ID NO: 1812 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

67. The AAV particle of claim 66, wherein the viral genome further comprises a filler sequence comprising the nucleotide sequence of SEQ ID NO: 1840 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto, optionally wherein the filler sequence is positioned 3’ to the polyA region and 5’ to the 3’ ITR.

68. The AAV particle of claim 62 or claim 63, wherein: (i) the 5’ ITR region comprises the nucleotide sequence of SEQ ID NO: 1811 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (ii) the promoter consists of the nucleotide sequence of SEQ ID NO: 1738 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iii) the intron / exon region comprises the nucleotide sequence of SEQ ID NO: 1816 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (v) the at least one miR122 binding site comprises a miR122 binding site series comprising SEQ ID NO: 1826 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) the polyA region comprises the nucleotide sequence of SEQ ID NO: 1828 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and / or (vii) the 3’ ITR region comprises SEQ ID NO: 1812 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

69. The AAV particle of claim 68, wherein the viral genome further comprises a filler sequence comprising the nucleotide sequence of SEQ ID NO: 1838 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, atleast 97%, at least 98%, or at least 99% identical) thereto, optionally wherein the filler sequence is positioned 3’ to the polyA region and 5’ to the 3’ ITR.

70. The AAV particle of claim 62 or claim 63, wherein: (i) the 5’ ITR region comprises the nucleotide sequence of SEQ ID NO: 1811 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (ii) the promoter consists of the nucleotide sequence of SEQ ID NO: 1740 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iii) the intron / exon region comprises the nucleotide sequence of SEQ ID NO: 1816 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (iv) the FXN-encoding sequence comprises the nucleotide sequence of SEQ ID NO: 1824 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (v) the at least one miR122 binding site comprises a miR122 binding site series comprising SEQ ID NO: 1826 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; (vi) the polyA region comprises the nucleotide sequence of SEQ ID NO: 1828 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and / or (vii) the 3’ ITR region comprises SEQ ID NO: 1812 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto.

71. The AAV particle of claim 70, wherein the viral genome further comprises a filler sequence comprising the nucleotide sequence of SEQ ID NO: 1839 or a sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto, optionally wherein the filler sequence is positioned 3’ to the polyA region and 5’ to the 3’ ITR.

72. The AAV particle of any one of claims 1-32, wherein the viral genome comprises: (a) the nucleotide sequence of SEQ ID NO: 1797 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, atleast 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1797; (b) the nucleotide sequence of SEQ ID NO: 1801 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1801; (c) the nucleotide sequence of SEQ ID NO: 1808 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1808; or (d) the nucleotide sequence of SEQ ID NO: 1809 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1809.

73. The AAV particle of any one of claims 9-15, wherein the viral genome comprises: (a) the nucleotide sequence of SEQ ID NO: 1797 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1797; (b) the nucleotide sequence of SEQ ID NO: 1801 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1801; (c) the nucleotide sequence of SEQ ID NO: 1808 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1808; or (d) the nucleotide sequence of SEQ ID NO: 1809 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1809.

74. The AAV particle of any one of claims 16-22, wherein the viral genome comprises: (a) the nucleotide sequence of SEQ ID NO: 1797 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, atleast 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1797; (b) the nucleotide sequence of SEQ ID NO: 1801 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1801; (c) the nucleotide sequence of SEQ ID NO: 1808 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1808; or (d) the nucleotide sequence of SEQ ID NO: 1809 or a nucleotide sequence that is at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) to the nucleotide sequence of SEQ ID NO: 1809.

75. An adeno-associated virus (AAV) particle comprising a viral genome comprising the nucleotide sequence of SEQ ID NO: 1797 and an AAV capsid variant comprising: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:

4.

76. An adeno-associated virus (AAV) particle comprising a viral genome comprising the nucleotide sequence of SEQ ID NO: 1797 and an AAV capsid variant comprising: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36; and / or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO:

36.

77. The AAV particle of any one of claims 1-76, wherein the viral genome is single-stranded.

78. A cell comprising the AAV particle of any one of claims 1-77, optionally wherein the cell is a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an Sf9 cell), or a bacterial cell.

79. A method of making an AAV particle of any one of claims 1-77, wherein the method comprises:(i) providing a cell comprising the viral genome comprising a frataxin (FXN)-encoding sequence and a nucleic acid encoding the AAV capsid variant; and (ii) incubating the cell under conditions suitable to encapsulate the viral genome in the AAV capsid variant; thereby making the AAV particle.

80. The method of claim 79, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 1797, or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO: 4; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 138-742 SEQ ID NO: 4; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 4 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 203-742 of SEQ ID NO:

4.

81. The method of claim 80, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 4, the amino acid sequence of positions 138-742 of SEQ ID NO: 4, and / or the amino acid sequence of positions 203-742 of SEQ ID NO:

4.

82. The method of claim 79, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 1797, or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO: 36;(ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 138-742 SEQ ID NO: 36; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 36 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 203-742 of SEQ ID NO:

36.

83. The method of claim 82, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 36, the amino acid sequence of positions 138-742 of SEQ ID NO: 36, and / or the amino acid sequence of positions 203-742 of SEQ ID NO:

36.

84. The method of claim 79, wherein the viral genome comprises the nucleotide sequence of SEQ ID NO: 1797, or a nucleotide sequence at least 90% identical (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical) thereto; and wherein the AAV capsid variant comprises: (i) a VP1 protein comprising the amino acid sequence of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to SEQ ID NO: 982; (ii) a VP2 protein comprising the amino acid sequence of positions 138-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 138-742 SEQ ID NO: 982; or (iii) a VP3 protein comprising the amino acid sequence of positions 203-742 of SEQ ID NO: 982 or an amino acid sequence having at least 90% identity (e.g., at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity) to positions 203-742 of SEQ ID NO:

982.

85. The method of claim 84, wherein the AAV capsid variant comprises the amino acid sequence of SEQ ID NO: 982, the amino acid sequence of positions 138-742 of SEQ ID NO: 982, and / or the amino acid sequence of positions 203-742 of SEQ ID NO:

982.

86. The method of any one of claims 79-85, further comprising, prior to step (i), introducing a first nucleic acid molecule comprising the viral genome into the cell.

87. The method of any one of claims 79-86, wherein the cell comprises a second nucleic acid molecule encoding the AAV capsid variant.

88. The method of claim 87, further comprising, prior to step (i), introducing the second nucleic acid molecule into the cell.

89. The metiiod of any one of claims 79-88, wherein the cell comprises a mammalian cell (e.g., an HEK293 cell), an insect cell (e.g., an Sf9 cell), or a bacterial cell.

90. A pharmaceutical composition comprising the AAV particle of any one of claims 1-77 and a pharmaceutically acceptable excipient.

91. A pharmaceutical composition comprising the AAV particle of any one of claims 5-22 and a pharmaceutically acceptable excipient.

92. A pharmaceutical composition comprising the AAV particle of any one of claims 9-15, 61, 64, 67, 70, and 75, and a pharmaceutically acceptable excipient.

93. A pharmaceutical composition comprising the AAV particle of any one of claims 16-22, 62, 65, 68, 71, and 76, and a pharmaceutically acceptable excipient.

94. A method of delivering a frataxin (FXN) protein to a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 90-93 or the AAV particle of any one of claims 1-77, thereby delivering the FXN protein.

95. The method of claim 94, wherein the subject has, has been diagnosed witii having, or is at risk of having a disorder associated with FXN deficiency.

96. The metiiod of claim 95, wherein the disorder associated with FXN deficiency is Friedreich’s Ataxia (FA).

97. A metiiod of treating a disorder associated witii frataxin (FXN) deficiency in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 90-93 or the AAV particle of any one of claims 1-77, thereby treating the disorder.

98. A method of treating a disorder associated with frataxin (FXN) deficiency in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of any one of claims 90-93 or the AAV particle of any one of claims 5-22, thereby treating the disorder.

99. A method of treating a disorder associated with frataxin (FXN) deficiency in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 92 or the AAV particle of any one of claims 9-15, 61, 64, 67, 70, and 75, thereby treating the disorder.

100. A method of treating a disorder associated with frataxin (FXN) deficiency in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 93 or the AAV particle of any one of claims 16-22, 62, 65, 68, 71, and 76, thereby treating the disorder.

101. The method of any one of claims 97-100, wherein the subject has, has been diagnosed with having, or is at risk of having a disorder associated with FXN deficiency.

102. The method of any one of claims 97-101, wherein the disorder is Friedreich’s Ataxia (FA).

103. The method of any one of claims 97-102, wherein the administration results in an increase in the subject’s FXN protein level as compared to baseline.

104. The method of any one of claims 97-103, wherein the treatment results in amelioration of at least one symptom of Friedreich’s Ataxia (FA).

105. The method of claim 104, wherein the at least one symptom of FA comprises impaired sensory functions, impaired motor function (e.g., ataxia and / or involuntary movements), fatigue, chronic pain, seizures, impaired speech, sleep disturbances, metabolic disorders (e.g., diabetes), and / or increased spasticity.

106. The method of any one of claims 100-105, wherein the treatment stabilizes, slows the progression of, or improves the subject’s FA as determined by the modified Friedreich Ataxia Rating Scale (mFARS), the Scale for the Assessment and Rating of Ataxia (SARA), and / or the International Cooperative Ataxia Rating Scale (ICARS).

107. The method of claim 106, wherein the treatment slows the subject’s progression of FA as measured by mFARS, SARA, and / or ICARS relative to an individual with the disorder associatedwith FXN deficiency who has not been administered the pharmaceutical composition or the AAV particle.

108. The method of any one of claims 97-107, wherein the subject is a human.

109. The method of any one of claims 97-108, wherein the AAV particle or the pharmaceutical composition is delivered to a cell or tissue of the CNS, optionally wherein the AAV particle or the pharmaceutical composition is delivered via intravenous administration.

110. The method of any one of claims 97-109, further comprising evaluating, e.g., measuring, the level of FXN expression, e.g., FXN gene, FXN mRNA, and / or FXN protein expression, in the subject, e.g., in a cell, tissue, or fluid, of the subject.

111. The method of claim 110, wherein the level of FXN protein expression is measured by an enzyme-linked immunosorbent assay (ELISA), a Western blot, an immunohistochemistry assay, or a frataxin biofluid assay.

112. The method of claim 110 or claim 111, wherein the cell or tissue is a cell or tissue of the central nervous system (CNS).

113. The method of claim 110 or claim 111, wherein the cell or tissue is a peripheral cell or tissue.

114. The method of any one of claims 97-113, wherein the administration results in an increase in: (i) the level of FXN protein or FXN gene expression in a cell, tissue, (e.g., a cell or tissue of the CNS, e.g., the cortex, striatum, thalamus, cerebellum, and / or brainstem), and / or fluid (e.g., CSF and / or serum), of the subject; and / or (ii) the level of viral genomes (VG) per cell in a CNS tissue (e.g., the cortex, striatum, thalamus, cerebellum, brainstem, and / or spinal cord) of the subject, optionally wherein the VG level is increased by greater than 50 VGs per cell, as compared to a peripheral tissue.

115. The method of any one of claims 97-114, further comprising administering to the subject at least one additional therapeutic agent and / or therapy.

116. The method of claim 115, wherein the at least one additional therapeutic agent and / or therapy comprises an agent and / or therapy for treating the disorder associated with FXN deficiency (e.g., Friedreich’s Ataxia).

117. The method of claim 115 or claim 116, wherein the at least one additional therapeutic agent and / or therapy comprises omaveloxolone or idebenone.

118. The method of any one of claims 97-117, further comprising administering an immunosuppressant to the subject.

119. The method of claim 118, wherein the immunosuppressant comprises a corticosteroid (e.g., prednisone, prednisolone, methylprednisolone, and / or dexamethasone), rapamycin, mycophenolate mofetil, tacrolimus, rituximab, and / or eculizumab hydroxychloroquine.

120. The pharmaceutical composition of any one of claims 90-93 or the AAV particle of any one of claims 1-77 for use in a method of treating a disorder according to any one of claims 100-119.

121. The pharmaceutical composition of any one of claims 90-93 or the AAV particle of any one of claims 1-77 for use in the treatment of a disorder associated with FXN deficiency in a subject, optionally wherein the disorder is Friedreich’s Ataxia.

122. The pharmaceutical composition or the AAV particle for use of claim 121, wherein the subject has, has been diagnosed with having, or is at risk of having Friedreich’s Ataxia.

123. Use of an effective amount of the pharmaceutical composition of any one of claims 90-93 or the AAV particle of any one of claims 1-77 in the manufacture of a medicament for the treatment of a disorder associated with FXN deficiency in a subject, optionally wherein the disorder is Friedreich’s Ataxia.

124. The use of claim 123, wherein the subject has, has been diagnosed with having, or is at risk of having Friedreich’s Ataxia.

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