US 11,851,694 B1
High fidelity in vitro transcription
David Mauger, Arlington, MA (US); Vladimir Presnyak, Manchester, NH (US); Amy E. Rabideau, Waltham, MA (US); and Iain Mcfadyen, Arlington, MA (US)
Assigned to ModernaTX, Inc., Cambridge, MA (US)
Filed by ModernaTX, Inc., Cambridge, MA (US)
Filed on Feb. 19, 2020, as Appl. No. 16/794,318.
Claims priority of provisional application 62/808,099, filed on Feb. 20, 2019.
Int. Cl. C12P 19/34 (2006.01); C12N 15/11 (2006.01)
CPC C12P 19/34 (2013.01) [C12N 15/11 (2013.01)] 15 Claims
 
1. A method of producing a messenger RNA (mRNA), the method comprising
(a) identifying a non-canonical terminator motif NNATCTGTTNN (SEQ ID NO: 50) in a DNA encoding a mRNA that encodes a polypeptide, wherein N at each position of the non-canonical terminator motif is any nucleotide selected from A, T, C, and G;
(b) producing a modified DNA comprising at least one codon substitution in the non-canonical terminator motif that preserves the amino acid sequence of the polypeptide; and
(c) producing a modified mRNA in an in vitro transcription reaction that comprises the modified DNA, nucleoside triphosphates, and a T7 RNA polymerase.