Rna polymerase variants
AssigneeModernaTX, Inc.
InventorsAmy E. Rabideau, Athanasios Dousis, Kanchana Ravichandran, Elissa Hobert
Life of this patent
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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.
Claims28
A ribonucleic acid (RNA) polymerase variant comprising an amino acid substitution, relative to wild-type RNA polymerase, that causes a loop structure of the RNA polymerase variant to undergo a conformational change to a helix structure as the RNA polymerase variant transitions from an initiation complex to an elongation complex.
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A ribonucleic acid (RNA) polymerase variant comprising:
- an additional C-terminal amino acid relative to wild-type RNA polymerase;
- a C terminus that comprises a FAFAXn (SEQ ID NO:
- 171) motif, wherein X is any amino acid and n is any integer greater than zero;
- a C terminus that comprises a FAFAG (SEQ ID NO:
- 329) motif;
- or
- a C terminus that comprises a XAFAXn motif, a FXFAXn motif, a FAXAXn motif, or a FAFXXn motif, wherein each X is any amino acid, and n is any integer greater than zero.
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A co-transcriptional capping method for ribonucleic acid (RNA) synthesis, the method comprising reacting a polynucleotide template with a RNA polymerase variant, optionally a T7 RNA polymerase variant, nucleoside triphosphates, and a cap analog under in vitro transcription reaction conditions to produce RNA transcript.
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Dependencies are inferred from the claim text.