Modified nucleosides, nucleotides, and nucleic acids, and uses thereof

Patent metadata

Field	Value
Publication number	US-9334328-B2
Application number	US-201313739212-A
Country	US
Kind code	B2
Filing date	Jan 11, 2013
Priority date	Oct 1, 2010
Publication date	May 10, 2016
Grant date	May 10, 2016

Abstract

The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.

First claim

What is claimed is:

1. A method of synthesizing a ribonucleic acid (RNA), comprising the steps of:
a) providing a complementary deoxyribonucleic acid (cDNA) that encodes a protein of interest;
b) selecting a nucleotide that is known to disrupt a binding of a major groove binding partner with a RNA, wherein the nucleotide has decreased binding affinity to the major groove binding partner selected from the group consisting of toll-like receptor (TLR) 3, TLR7, TLR8, retinoic acid-inducible gene I (RIG-I), melanoma differentiation-associated gene 5 (MDA5) and laboratory of genetics and physiology 2 (LGP2), and wherein the nucleotide comprises a modification on the major groove face of the nucleobase where an atom of the major groove face of the nucleobase is replaced or substituted with an alkyl group; and
c) contacting the provided cDNA and the selected nucleotide with an RNA polymerase, under conditions such that the RNA is synthesized.

2. The method of claim 1 , wherein the ribonucleic acid (RNA) is a messenger RNA (mRNA).

3. The method of claim 1 , wherein the major groove binding partner is TLR3.

4. The method of claim 1 , wherein the major groove binding partner is TLR7.

5. The method of claim 1 , wherein the major groove binding partner is TLR8.

6. The method of claim 1 , wherein the major groove binding partner is RIG-I.

7. The method of claim 1 , wherein the major groove binding partner is MDA5.

8. The method of claim 1 , wherein the major groove binding partner is LGP2.

9. The method of claim 1 , wherein the nucleobase comprising a modification on the major groove face is a pyrimidine nucleobase.

10. The method of claim 2 , wherein the nucleobase comprising a modification on the major groove face is a pyrimidine nucleobase.

Assignees

• Moderna Therapeutics Inc

Inventors

• Schrum Jason P
• Siddiqi Suhaib
• Ejebe Kenechi

Classifications

• C12P21/00

  Preparation of peptides or proteins (single cell protein C12N1/00) · CPC title

• C07H21/02 Primary

  with ribosyl as saccharide radical · CPC title

• C12N15/1138

  against receptors or cell surface proteins · CPC title

• C07H19/10

  with the saccharide radical esterified by phosphoric or polyphosphoric acids · CPC title

• C12N2310/335

  Modified T or U · CPC title