IP Library › Granted Patent US 12,312,610
Granted Patent B2
US 12,312,610 · App. 18/462,868 · Granted May 27, 2025

Vaccinia capping enzyme compositions and methods

Inventors: Saulius Vainauskas (Newburyport, MA); Siu-hong Chan (Ipswich, MA); Christopher H. Taron (Essex, MA)
Assignee: New England Biolabs, Inc.
C12N9/1241C12N15/815C07K2319/00C12R2001/84C12Y207/0705
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Quick Facts
Patent No.
US 12,312,610
App. No.
18/462,868
Granted
May 27, 2025
Kind
B2
Abstract

The present disclosure relates, according to some embodiments, to compositions, methods, and/or kits for producing vaccinia capping enzyme. For example, active, heterodimers of vaccinia capping enzyme may be produced as fusions comprising D1 and D12 subunits. Vaccinia capping enzyme fusion proteins may further comprise a linker.

Claims (22)

1. A vaccinia capping enzyme fusion transcript comprising, in a 5′ to 3′ direction: (a) a sequence encoding a D1 subunit, (b) a sequence encoding a linker, and (c) a sequence encoding a D12 subunit, wherein the linker has at least 90% identity to SEQ ID NO: 3.

2. A composition comprising a vaccinia capping enzyme fusion transcript, wherein the vaccinia capping enzyme fusion transcript comprises, in a 5′ to 3′ direction:

(a) a sequence encoding a D1 subunit;

(b) a sequence encoding a linker; and

(c) a sequence encoding a D12 subunit, wherein the linker has at least 90% identity to SEQ ID NO: 3.

3. A composition according to claim 2 , wherein the D1 subunit has an amino acid sequence having at least 90% identity to positions 24 to 867 of SEQ ID NO: 1.

4. A composition according to claim 2 , wherein the D1 subunit has an amino acid sequence having at least 90% identity to SEQ ID NO: 1.

5. A composition according to claim 2 , wherein the D12 subunit has an amino acid sequence having at least 90% identity to SEQ ID NO: 2.

6. A composition according to claim 2 , wherein the vaccinia capping enzyme fusion transcript further comprises (d) a cap.

7. A cell comprising the composition according to claim 2 .

8. A cell according to claim 7 , further comprising equimolar amounts of the D1 subunit and the D12 subunit.

9. A cell according to claim 7 , further comprising an active vaccinia capping enzyme fusion protein.

10. A cell according to claim 8 , wherein the vaccinia capping enzyme fusion protein has an amino acid sequence at least 90% identical to SEQ ID NO: 4 or at least 90% identical to SEQ ID NO: 6 or at least 90% identical to SEQ ID NO: 8.

11. A method for producing an active vaccinia capping enzyme comprising:

contacting

(a) a vaccinia capping enzyme fusion transcript comprising, in a 5′ to 3′ orientation,

(i) a sequence encoding D1, (ii) a sequence encoding a linker having at least 90% identity to SEQ ID NO: 3, and

(iii) a sequence encoding D12; with

(b) an expression system.

12. A method according to claim 11 , wherein the expression system is a yeast expression system.

13. A method according to claim 12 , wherein the yeast is Kluyveromyces lactis or Pichia pastoris.

14. A cell according to claim 7 , wherein the cell is a Kluyveromyces lactis cell or a Pichia pastoris cell.

Assignments (2)
SECURITY INTEREST Recorded May 29, 2026
From: NEW ENGLAND BIOLABS, INC.
To: BANK OF AMERICA, N.A., AS ADMINISTRATIVE AGENT
Reel/Frame 074795/0566 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 7, 2023
From: VAINAUSKAS, SAULIUS; CHAN, SIU-HONG; TARON, CHRISTOPHER H.
To: NEW ENGLAND BIOLABS, INC.
Reel/Frame 064839/0209 →
Continuity (3)
Division 17348127 · Jun 15, 2021
Provisional Application 63042939 · Jun 23, 2020
Related Publication 20230416701A1 · Dec 28, 2023
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