Reducing intron retention

Patent metadata

Field	Value
Publication number	US-11891605-B2
Application number	US-202217846794-A
Country	US
Kind code	B2
Filing date	Jun 22, 2022
Priority date	Jun 16, 2014
Publication date	Feb 6, 2024
Grant date	Feb 6, 2024

Abstract

Disclosed herein are methods, compositions, polynucleic acid polymers, assays, and kits for inducing processing of a partially processed mRNA transcript to remove a retained intron to produce a fully processed mRNA transcript that encodes a full-length functional form of a protein. Also described herein are methods and compositions for treating a disease or condition characterized by impaired production of a full-length functional form of a protein or for treating a disease or condition characterized by a defective splicing in a subject.

First claim

What is claimed is:

1. A method of making a polynucleic acid for inducing processing of a partially processed mRNA transcript to facilitate removal of a retained intron to produce a fully processed mRNA transcript that encodes a functional form of a protein, the method comprising:
synthesizing the polynucleic acid, wherein the polynucleic acid is from about 10 to about 50 nucleotides in length and comprises a sequence that is complementary to at least 10 contiguous bases of a wild-type target sequence of the partially processed mRNA transcript, wherein the partially processed mRNA transcript is capable of encoding the functional form of a protein and comprises at least one retained intron, wherein the polynucleic acid induces splicing out of the retained intron from the partially processed mRNA transcript.

2. The method of claim 1 , wherein the polynucleic acid hybridizes to the wild-type target sequence of the partially processed mRNA transcript.

3. The method of claim 1 , wherein the wild-type target sequence is a binding motif that forms a hairpin structure.

4. The method of claim 1 , wherein the wild-type target sequence is between two G quadruplexes of a partially processed mRNA transcript.

5. The method of claim 1 , wherein the retained intron consists of a canonical intronic sequence between two canonical exons of the partially processed mRNA transcript.

6. The method of claim 1 , wherein the wild-type target sequence does not form a G quadruplex.

7. The method of claim 1 , wherein the intronic splicing regulatory element comprises a first CCC motif.

8. The method of claim 1 , wherein the polynucleic acid is from 10 to 30 nucleotides in length.

9. The method of claim 1 , wherein the sequence of the polynucleic acid is at least 90% complementary to the wild-type target sequence of the partially processed mRNA transcript.

10. The method of claim 1 , wherein the polynucleic acid is modified at a nucleoside moiety, at a phosphate moiety, at a 5′ terminus, at a 3′ terminus, or a combination thereof.

11. The method of claim 10 , wherein the polynucleic acid comprises an artificial nucleotide.

12. The method of claim 11 , wherein the artificial nucleotide is selected from the group consisting of 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), 2′-O—N-methylacetamido (2′-O-NMA), a locked nucleic acid (LNA), an ethylene nucleic acid (ENA), a peptide nucleic acid (PNA), a 1′,5′-anhydrohexitol nucleic acid (HNA), a morpholino, a methylphosphonate nucleotide, a thiolphosphonate nucleotide, and a 2′-fluoro N3-P5′-phosphoramidite.

13. The method of claim 1 , wherein the method further comprises attaching the polynucleic acid to a delivery vehicle.

14. The method of claim 13 , wherein the delivery vehicle comprises a cell penetrating peptide or a peptide-based nanoparticle.

15. The method of claim 1 , wherein synthesizing comprises chemically synthesizing the polynucleic acid.

16. The method of claim 1 , wherein synthesizing comprises enzymatic ligation.

17. The method of claim 1 , wherein synthesizing comprises expressing the polynucleic acid from a vector.

Assignees

• Univ Southampton

Inventors

• Vorechovsky Igor
• Kralovicova Jana

Classifications

• C12N15/113 Primary

  Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)} · CPC title

• C12N15/111

  General methods applicable to biologically active non-coding nucleic acids · CPC title

• C12Q1/6883

  for diseases caused by alterations of genetic material · CPC title

• C12N2310/11

  Antisense · CPC title

• C12N2310/113

  targeting other non-coding nucleic acids, e.g. antagomirs · CPC title