US 12,479,880 B2
Enzymatic RNA capping method
G. B. Robb, Somerville, MA (US); Siu-hong Chan, Ipswich, MA (US); and Bijoyita Roy, Medford, MA (US)
Assigned to New England Biolabs, Inc., Ipswich, MA (US)
Filed by New England Biolabs, Inc., Ipswich, MA (US)
Filed on Aug. 5, 2024, as Appl. No. 18/794,874.
Application 18/794,874 is a division of application No. 17/001,236, filed on Aug. 24, 2020, granted, now 12,398,173.
Application 17/001,236 is a continuation of application No. PCT/US2020/047521, filed on Aug. 21, 2020.
Application 17/001,236 is a continuation of application No. PCT/US2020/047533, filed on Aug. 21, 2020.
Claims priority of provisional application 62/890,821, filed on Aug. 23, 2019.
Prior Publication US 2024/0383939 A1, Nov. 21, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C07H 19/20 (2006.01); C07H 21/04 (2006.01); C12P 19/34 (2006.01); C12Q 1/6816 (2018.01)
CPC C07H 19/20 (2013.01) [C07H 21/04 (2013.01); C12P 19/34 (2013.01); C12Q 1/6816 (2013.01)] 23 Claims
OG exemplary drawing
 
1. A composition comprising:
(i) a polynucleotide;
(ii) a single-chain RNA capping enzyme that has RNA triphosphatase (TPase), guanylyltransferase (GTase) and guanine-N7 methyltransferase (N7 MTase) activities and comprises an amino acid sequence at least 90% identical to (a) SEQ ID NO:2, (b) SEQ ID NO:3, and (c) SEQ ID NO:4;
(iii) guanosine triphosphate (GTP);
(iv) a buffering agent; and
(v) a methyl group donor,
wherein the amino acid sequence further has at least 85% identity to (A) SEQ ID NO:7, (B) SEQ ID NO: 8, (C) SEQ ID NO:9, and/or (D) SEQ ID NO:10.