IP Library › Granted Patent US 12,195,778
Granted Patent B2
US 12,195,778 · App. 18/448,856 · Granted Jan 14, 2025

RNA polymerase variants

Inventors: Amy E. Rabideau (Waltham, MA); Athanasios Dousis (Somerville, MA); Kanchana Ravichandran (Cambridge, MA); Elissa Hobert (Cambridge, MA)
Assignee: ModernaTX, Inc.
C12P19/34C12N9/1247C12Y207/11023
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Quick Facts
Patent No.
US 12,195,778
App. No.
18/448,856
Granted
Jan 14, 2025
Kind
B2
Abstract

The present disclosure provides, in some aspects, variant RNA polymerases, the use of which increases transcription efficiency while reducing the number of double-stranded RNA contaminates and run-on transcripts produced during an in vitro transcription reaction.

Claims (12)

1. A method of performing an in vitro transcription (IVT) reaction, comprising combining a deoxyribonucleic acid (DNA) template with a T7 ribonucleic acid (RNA) polymerase variant, nucleoside triphosphates, and buffer under conditions that result in the production of to produce an RNA transcript, wherein the T7 RNA polymerase variant comprises an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 1 modified to comprise an amino acid substitution at position.

2. The method of claim 1 , wherein the amino acid substitution is selected from the group consisting of alanine, isoleucine, leucine, methionine, lysine, glutamine, and glutamate.

3. The method of claim 2 , wherein the amino acid substitution is alanine.

4. The method of claim 1 , wherein the T7 RNA polymerase variant comprises an amino acid sequence having at least 95% identity to SEQ ID NO:1.

5. The method of claim 1 , wherein the T7 RNA polymerase variant comprises an amino acid sequence having at least 98% identity to SEQ ID NO:1.

6. The method of claim 1 , wherein the T7 RNA polymerase comprises an additional C-terminal amino acid.

7. The method of claim 6 , wherein the additional C-terminal amino acid comprises glycine (G).

8. The method of claim 1 , wherein the T7 RNA polymerase comprises two additional C-terminal amino acids.

9. The method of claim 8 , wherein the two additional C-terminal amino acids comprise the same type of amino acid.

10. The method of claim 8 , wherein the two additional C-terminal amino acids comprise two different types of amino acids.

11. The method of claim 1 , wherein the amino acid substitution at position S43 is alanine (S43A).

12. The method of claim 11 , wherein the RNA polymerase variant comprises the amino acid sequence of SEQ ID NO: 2.

Assignments (2)
SECURITY INTEREST Recorded Nov 19, 2025
From: MODERNATX, INC.
To: ARES CAPITAL CORPORATION, AS AGENT
Reel/Frame 073634/0354 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 25, 2024
From: RABIDEAU, AMY E.; DOUSIS, ATHANASIOS; RAVICHANDRAN, KANCHANA; HOBERT, ELISSA
To: MODERNATX, INC.
Reel/Frame 066249/0351 →
Continuity (9)
Continuation 17350662 · Jun 17, 2021
Continuation 16657122 · Oct 18, 2019
Division 16432541 · Jun 5, 2019
Continuation PCTUS2018046989 · Aug 17, 2018
Provisional Application 62677527 · May 29, 2018
Provisional Application 62638684 · Mar 5, 2018
Provisional Application 62628484 · Feb 9, 2018
Provisional Application 62547677 · Aug 18, 2017
Related Publication 20240102066A1 · Mar 28, 2024
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