Modified nucleosides, nucleotides, and nucleic acids, and uses thereof
View Patent ↗The present disclosure provides modified nucleosides, nucleotides, and nucleic acids, and methods of using thereof.
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.