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. A method of inhibiting nonsense-mediated decay (NMD) of mRNA in a gene-specific manner in a eukaryotic cell, comprising: contacting (a) a eukaryotic cell that comprises (i) a nucleic acid that contains a disease-causing premature termination codon (PTC) or a naturally-occurring premature termination codon (PTC), (ii) mRNA encoded by the nucleic acid, and (iii) an exon-exon junction in the mRNA that is downstream of a disease-causing or naturally occurring premature termination codon (PTC) and is bound by exon junction complex (EJC) that identifies the mRNA for nonsense-mediated decay (NMD), with (b) an antisense oligonucleotide (ASO) that hybridizes to a region of the mRNA that is from about 1 to about 50 nucleotides upstream of the exon-exon junction under conditions wherein the ASO enters the cell in sufficient quantity to inhibit deposition of EJC at the exon-exon junction of the mRNA that contains the PTC.
2. The method of claim 1 , wherein the disease-causing PTC results from a mutation.
3. The method of claim 1 , wherein the ASO is specific to a region of the nucleic acid that is from about 20 to about 24 nucleotides upstream of an exon-exon junction.
4. The method of claim 3 , wherein the ASO is no less than 14 nucleotides in length.
5. The method of claim 1 , wherein the nucleic acid is a HBB allele that contains a nonsense mutation.
6. The method of claim 5 , wherein the HBB allele is CM810001, CM880039, CM034660, CM880040, or CM900122.
7. The method of claim 5 , wherein the ASO is selected from SEQ ID NOs: 1-19.
8. The method of claim 1 , wherein the eukaryotic cell is in an individual.
9. The method of claim 8 , wherein the individual is a human.
10. The method of claim 1 , wherein the eukaryotic cell is ex vivo.
11. The method of claim 1 , wherein the nucleic acid is a HBB allele that contains a nonsense mutation and a disease associated with the PTC is β-thalassemia; the nucleic acid is a MECP2 allele that contains a nonsense mutation and a disease associated with the PTC is Rett syndrome; the nucleic acid is a CFTR allele that contains a nonsense mutation and a disease associated with the PTC is cystic fibrosis; the nucleic acid is a DMD allele that contains a nonsense mutation and a disease associated with the PTC is Duchenne muscular dystrophy; the nucleic acid is a BMD allele that contains a nonsense mutation and a disease associated with the PTC is Becker muscular dystrophy; or the nucleic acid is a IDUA allele that contains a nonsense mutation and a disease associated with the PTC is Mucopolysaccharidosis type 1-Hurler.