RNA polymerase variants

Patent metadata

Field	Value
Publication number	US-11767548-B2
Application number	US-202117350662-A
Country	US
Kind code	B2
Filing date	Jun 17, 2021
Priority date	Aug 18, 2017
Publication date	Sep 26, 2023
Grant date	Sep 26, 2023

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.

First claim

What is claimed is:

1. A T7 ribonucleic acid (RNA) polymerase variant comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 1 modified to comprise an amino acid substitution at position S43, wherein the T7 RNA polymerase variant has RNA polymerase activity.

2. A composition or kit comprising the RNA polymerase variant of claim 1 and optionally an in vitro transcription (IVT) reagent.

3. The T7 RNA polymerase variant of claim 1 , wherein the amino acid substitution at position S43 is selected from the group consisting of alanine, isoleucine, leucine, methionine, lysine, glutamine, glutamate, and arginine.

4. The T7 RNA polymerase variant of claim 3 , wherein the amino acid substitution at position S43 is alanine (S43A).

5. The T7 RNA polymerase variant 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 T7 RNA polymerase variant of claim 1 , wherein the RNA polymerase variant comprises the amino acid sequence of SEQ ID NO: 2.

7. A T7 ribonucleic acid (RNA) polymerase variant comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 1 modified to comprise an alanine at position 43, wherein the T7 RNA polymerase variant has RNA polymerase activity.

8. The T7 RNA polymerase variant of claim 7 comprising the amino acid sequence of SEQ ID NO: 2.

Assignees

• Modernatx Inc

Inventors

• Rabideau Amy E
• Dousis Athanasios
• Ravichandran Kanchana
• Hobert Elissa

Classifications

• C12P19/34 Primary

  Polynucleotides, e.g. nucleic acids, oligoribonucleotides · CPC title

• C12N9/1247 Primary

  DNA-directed RNA polymerase (2.7.7.6) · CPC title

• C12Y207/11023

  [RNA-polymerase-subunit] kinase (2.7.11.23) · CPC title

• C12Y207/07006

  DNA-directed RNA polymerase (2.7.7.6) · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title