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US 10,465,190 B1Granted
In vitro transcription methods and constructs
AssigneeModernaTX, Inc.
InventorsJesse Chen, Athanasios Dousis
Life of this patent
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Abstract
The present disclosure provides, in some aspects, in vitro transcription systems (including, for example, nucleic acid constructs and polymerases), the use of which increases transcription efficiency while reducing the amount of truncated single-stranded ribonucleic acid transcript produced during an in vitro transcription reaction.
Claims14
A method of producing a ribonucleic acid (RNA) of interest, the method comprising:
- (a) performing an in vitro transcription reaction using an initial amount of an in vitro transcription construct that comprises a modified T7 promoter operably linked to a nucleic acid that comprises a 5′ untranslated region (UTR) and an open reading frame encoding the RNA of interest, wherein the modified T7 promoter comprises a point mutation at position −14, −15, and/or −16, relative to a wild-type T7 promoter that comprises the nucleotide sequence of SEQ ID NO:
- 18, and the 5′ UTR comprises the T7 bacteriophage Gene 9 sequence of SEQ ID NO:
- 12;
- and
- (b) producing the RNA of interest.
2 more elements
Dependencies are inferred from the claim text.