US 12,305,210 B2
In vitro transcription technologies
Irena Rabe, Mainz (DE); Maximilian Buff, Hofheim (DE); Thomas Ziegenhals, Undenheim (DE); Johanna Drögemüller, Weiterstadt (DE); Andreas Kuhn, Mainz (DE); Stephanie Fesser, Mannheim (DE); Rodney Gene Combs, Wildwood, MO (US); Nicole Star Eschmann, St Peters, MO (US); Jennifer Ann Schoborg Romine, St. Louis, MO (US); and Jenna Kathryn Williamson, Chesterfield, MO (US)
Assigned to BioNTech SE, Mainz (DE)
Filed by BioNTech SE, Mainz (DE)
Filed on Aug. 23, 2022, as Appl. No. 17/893,742.
Claims priority of provisional application 63/236,660, filed on Aug. 24, 2021.
Claims priority of application No. PCT/EP2021/074139 (WO), filed on Sep. 1, 2021.
Prior Publication US 2023/0183769 A1, Jun. 15, 2023
Int. Cl. C12P 19/34 (2006.01); C12N 9/12 (2006.01)
CPC C12P 19/34 (2013.01) [C12N 9/1247 (2013.01)] 18 Claims
OG exemplary drawing
 
1. A method of producing a ribonucleic acid (RNA) molecule through in vitro transcription, the method comprising creating a reaction mixture under reaction conditions to form the RNA molecule, the reaction mixture comprising a nucleic acid polymerase, a nucleic acid template, and:
a molar ratio a of total cytidine triphosphate (CTP) and/or one or more CTP analog(s) to total guanosine triphosphate (GTP) and/or one or more GTP analog(s); and/or
a molar ratio b of total CTP and/or one or more CTP analog(s) to total uridine triphosphate (UTP) and/or one or more UTP analog(s); and/or
a molar ratio c of total CTP and/or one or more CTP analog(s) to total adenosine triphosphate (ATP) and/or one or more ATP analog(s),
wherein:
a is at least 1.25; and/or
b is at least 1.25; and/or
c is at least 1.10,
so that the molar ratio of total CTP to GTP to UTP to ATP is not 1:1:1:1, and further wherein;
the method is independent of the sequence of the RNA molecule.