# US 10,946,035 B2 · Production of stable non-polyadenylated RNAs

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- Number: US 10,946,035 B2
- Document ID: US10946035B2
- Status: Granted
- Kind: B2
- Application number: 15/664,017
- Filed: 2017-07-31
- Published: 2018-05-24
- Granted: 2021-03-16
- Earliest claimed: 2012-10-16
- Assignees: Massachusetts Institute of Technology
- Applicants: Massachusetts Institute of Technology
- Inventors: Jeremy E. Wilusz; Phillip A. Sharp
- Web page: https://patentgrep.com/patent/US10946035B2

## CPC

- A61K 31/7088: Compounds having three or more nucleosides or nucleotides
- A61K 31/7105: Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
- A61K 48/00: Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61P 9/10: for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
- A61P 11/00: Drugs for disorders of the respiratory system
- A61P 17/00: Drugs for dermatological disorders
- A61P 21/02: Muscle relaxants, e.g. for tetanus or cramps
- A61P 25/28: for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
- A61P 35/00: Antineoplastic agents
- A61P 37/02: Immunomodulators
- C12N 15/113: 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)}
- C12N 15/67: General methods for enhancing the expression
- C12N 2310/15: Nucleic acids forming more than 2 strands, e.g. TFOs
- C12N 2310/3519: Fusion with another nucleic acid
- C12N 2320/52: modulating the physical stability, e.g. GC-content
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The invention relates in aspects to hybrid RNAs lacking a poly-A tail and nucleic acid vectors for expressing the RNA. The hybrid RNAs in some instances have a stabilizing triple helical structure. Related methods for expressing RNA in vivo and in vitro are also disclosed.

## Claims

### Claim 1

A method for expressing an RNA lacking a poly-A tail, comprising: expressing in a cell an isolated nucleic acid comprising an RNA having a 3′ heterologous terminal sequence and lacking a poly A tail, wherein the heterologous terminal sequence comprises an RNA stabilizing terminal sequence that has a triple helix conformation, and wherein the immunogenic protein is a viral protein or a cancer protein.

### Claim 2 (depends on 1)

The method of claim 1, wherein the isolated nucleic acid is expressed in vivo.

### Claim 3 (depends on 1)

The method of claim 1, wherein the isolated nucleic acid is expressed ex vivo.

### Claim 4 (depends on 1)

The method of claim 1, wherein the RNA encodes a protein.

### Claim 5 (depends on 4)

The method of claim 4, wherein the protein is an immunogenic protein.

### Claim 6

A method for vaccinating a subject, comprising:
administering to a subject a hybrid nucleic acid comprising an RNA molecule, lacking a poly-A tail, linked to a heterologous RNA stabilizing terminal sequence, wherein the RNA molecule encodes an immunogenic protein, in an effective amount to elicit an adaptive immune response to the immunogenic protein, wherein the heterologous RNA stabilizing terminal sequence has a triple helix conformation.

### Claim 7 (depends on 6)

The method of claim 6, wherein the immunogenic protein is a fusion protein encompassing an antigenic protein or fragment thereof fused with a carrier peptide.

### Claim 8 (depends on 6)

The method of claim 6, wherein the immunogenic protein is a viral or bacterial antigen.

### Claim 9 (depends on 6)

The method of claim 6, wherein the immunogenic protein is a cancer antigen.

### Claim 10 (depends on 6)

The method of claim 6, wherein the immunogenic protein is an epitope.

### Claim 11 (depends on 10)

The method of claim 10, wherein the epitope is a MHC Class I epitope, a MHC Class II epitope, a B cell epitope or a T cell epitope.