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

A method of inducing in vivo translation of a recombinant polypeptide in a mammalian subject in need thereof, comprising administering to the mammalian subject a synthetic messenger RNA (mRNA) in an effective amount such that the synthetic mRNA is localized into a cell population of the mammalian subject and the recombinant polypeptide is translated in the cell population from the synthetic mRNA,

• wherein the synthetic mRNA is prepared by a synthetic method comprising the steps of:
• a) providing a complementary deoxyribonucleic acid (cDNA) encoding a pharmaceutical protein of interest;
• b) selecting a nucleotide that disrupts a binding of a major groove binding partner with an mRNA, 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 an RNA transcript encoding the recombinant polypeptide is synthesized.