Enzymes and polymerases for the synthesis of RNA

摘要

The invention relates to compositions and methods for the design, evolution, preparation, and/or manufacture of enzymes for use with polynucleotides, primary transcripts and mmRNA molecules.

主权项

1. A chimeric enzyme for synthesizing a capped RNA molecule, said chimeric enzyme comprising at least one capping enzyme and at least one nucleic acid polymerase, wherein said capped RNA molecule comprises at least one chemical modification, and wherein the at least one capping enzyme comprises a Vaccinia capping enzyme catalytic subunit (D1) and a Vaccinia capping enzyme stimulation subunit (D12), and where the at least one nucleic acid polymerase is T7 RNA polymerase (T7) in one of the following combinations:
a) a single polypeptide comprising D1 linked to D12 linked to T7; b) a single polypeptide comprising D12 linked to D1 linked to T7; c) a single polypeptide comprising T7 linked to D1 linked to D12; or d) a single polypeptide comprising T7 linked to D12 linked to D1.