MORGADO PEREIRA DE ALMEIDA LUÍS FERNANDO (PT)
CENTRO DE NEUROCIENCIAS E BIOLOGIA CELULAR (PT)
| AMENDED CLAIMS received by the International Bureau on 13 July 2020 (13.07.2020) 1. A ribonucleic add (RNA) molecule comprising : an antisense ribonucleotide sequence base-paired to a substantially complementary sense ribonucleotide sequence; wherein each ribonucleotide of the antisense RNA sequence is complementary to a corresponding ribonucleotide of a mutant human ataxin-3 comprising a single nucleotide polymorphism in linkage disequilibrium with a Machado-Joseph disease (MJD) allele of the mutant human ataxin-3 gene, and wherein the ribonucleotide of the antisense RNA sequence that is complementary to the single nucleotide polymorphism of the mutant human ataxin-3 mRNA is 10 ribonucleotides apart from the ribonucleotide at the 5' end of the antisense RNA sequence. 2. The RNA molecule of claim 1, wherein the base-paired sense ribonucleotide sequence is not fully complementary to the antisense ribonucleotide sequence. 3. The RNA molecule of claim 1, wherein the antisense ribonucleotide sequence is at least 90% complementary to the sense ribonucleotide sequence. 4. The RNA molecule of claim 1, wherein the antisense ribonucleotide sequence is complementary to SEQ ID NO. 1 or 13. 5. The RNA molecule of claim 1, wherein the antisense ribonucleotide sequence is SEQ ID NO. 2, 3, 4, 5, 6, 14, 15, 16, 17 or 18. 6. The RNA molecule of claim 1, wherein the antisense ribonucleotide sequence is SEQ ID NO. 2. 7. The RNA molecule of claim 1, wherein the ribonucleotides at the 5' and 3' ends of the antisense RNA sequence are at least 17 ribonucleotides apart. 8. The RNA molecule of claim 1, wherein the ribonucleotides at the 5' and 3' ends of the antisense RNA sequence are 17-21 ribonucleotides apart. 9. The RNA molecule of claim 1, wherein the RNA molecule is a single RNA molecule. 10. The RNA molecule of claim 7, wherein the RNA molecule is a miRNA. 11. The RNA molecule of claim 1, wherein the single nucleotide polymorphism (SNP) comprises an SNP of a rs1048755 (exon 8) or rs12895357 (exon 10) allele of the human ataxin-3 gene. 12. The RNA molecule of claim 1, wherein the RNA molecule is therapeutically effective at selectively silencing the expression of the Machado-Joseph disease (MJD) allele of the mutant human ataxin-3 gene but not a wild type human ataxin-3 allele. 13. The RNA molecule of claim 1 comprising a miRNA scaffold derived from miR-155, wherein the antisense ribonucleotide is SEQ ID NO. 2 and the sense ribonucleotide is SEQ ID NO. 1. 14. An adeno-associated viral vector comprising an isolated DNA sequence operably linked to a promoter, wherein the DNA sequence encodes the RNA molecule of claim 1. 15. A method for selectively silencing the expression of a mutant human ataxin-3 allele having a single nucleotide polymorphism in linkage disequilibrium with a Machado- Joseph disease (MJD) allele of the mutant human ataxin-3 comprising administering the adeno-associated viral vector of claim 12 to a subject in need thereof. 16. The method of claim 13, wherein the single nucleotide polymorphism (SNP) comprises an SNP of a rs1048755 (exon 8) or rs12295357 (exon 10) allele of the mutant human ataxin-3 gene. 17. The method of claim 14, wherein the adeno-associated viral vector is administered systemically, intravenously, intratumorally, orally, intranasally, intra peritoneal ly, intramuscularly, intravertebrally, intracerebrally, intracerebroventriculally, intracisternally, intratheca I ly, intraocularly, intracardiacally, intradermally, or subcutaneously, preferably intravenously, intracisternally, intrathecally or, in situ, by intracerebral administration. |