Reducing nonsense-mediated mRNA decay
The present disclosure relates to compositions and methods for inhibiting nonsense-mediated mRNA decay in a gene-specific manner, for example in the treatment of diseases or disorders caused by nonsense mutations.
1. An antisense oligonucleotide (ASO) that is complementary to a region of mRNA that is transcribed in eukaryotic cells and contains a disease-causing premature termination codon (PTC) derived from a CFTR gene containing a nonsense mutation, wherein the exon-exon junction is located downstream of the PTC, the disease is cystic fibrosis and the region is from about 1 to about 50 nucleotides upstream of the exon-exon junction.
2. An antisense oligonucleotide (ASO) specific to a region of mRNA that is transcribed in a eukaryotic cell, wherein the region of the mRNA is from about 20 to about 24 nucleotides upstream of an exon-exon junction (from about −20 to about −24 nucleotides) that is downstream of a disease-causing premature termination codon (PTC) in the mRNA, wherein the disease is cystic fibrosis and the PTC is derived from a CFTR gene that contains codon change TGGa-TGA, which results in nonsense mutation TRP-Ter and to which deposition of exon junction complexes (EJC) marks the mRNA for nonsense-mediated decay, wherein the ASO is at least 14 nucleotides.
3. An antisense oligonucleotide (ASO) that is complementary to a region of an mRNA that is transcribed in eukaryotic cells and contains a disease-causing premature termination codon (PTC) derived from a CFTR gene containing a nonsense mutation and an exon-exon junction located downstream from the PTC, wherein the disease is cystic fibrosis and the region is from about 1 to about 50 nucleotides upstream of the exon-exon junction, wherein the CFTR gene containing a nonsense mutation is CM900061.
4. The ASO of claim 1 , wherein the ASO comprises SEQ ID NO: 107, SEQ ID NO: 83 or SEQ ID NO: 65.
5. The ASO of claim 3 , wherein the ASO comprises SEQ ID NO: 107, SEQ ID NO: 83 or SEQ ID NO: 65.
6. The ASO of claim 5 , wherein the region of the mRNA is from about 20 to about 24 nucleotides upstream of the exon-exon junction (from about 20 to about 24 nucleotides).
7. The ASO of claim 5 , wherein the region of the mRNA is an Exon Junction Complex-protected area or a region adjacent to an Exon-Junction Complex-protected area.
8. The ASO of claim 4 which is at least 95% complementary to the region of the mRNA.
9. The ASO of claim 4 which is from 14 nucleotides to 20 nucleotides.
10. The ASO of claim 2 , wherein the ASO hybridizes and remains bound to the region under physiological conditions.
11. The ASO of claim 2 , wherein the allele is a CFTR allele and the ASO comprises SEQ ID NO: 107, SEQ ID NO: 83 or SEQ ID NO: 65.
12. An antisense oligonucleotide (ASO) of claim 1 , wherein the mRNA is expressed from a CFTR gene that contains the codon change TGGa-TGA at codon 1282 and the amino acid change is Trp-Ter.
13. An antisense oligonucleotide (ASO) that is complementary to a region of an mRNA that (a) is transcribed in eukaryotic cells and (b) contains (1) a disease-causing premature termination codon (PTC) derived from a CFTR gene that contains a nonsense mutation or a naturally-occurring premature termination codon (PTC) and (2) an exon-exon junction located downstream from the PTC, wherein the disease is cystic fibrosis and the region is from about 1 to about 50 nucleotides upstream of the exon-exon junction.
14. An antisense oligonucleotide (ASO) of claim 13 , wherein the region of the mRNA is 1 to 49 nucleotides upstream of the exon-exon junction.
15. An antisense oligonucleotide (ASO) of claim 14 , wherein the region of the mRNA is 1 to 40 nucleotides upstream of the exon-exon junction.