Novel Single-Subunit Rna Polymerase

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This patent discloses a novel DNA-directed single-subunit RNA polymerase (ssRNAP), named RNAP_E, derived from a viral source in elephant feces. RNAP_E offers several performance advantages over classic phage RNA polymerases (like T7), including higher thermo-stability, faster RNA synthesis, distinct promoter specificity, and tolerance to non-canonical nucleotides. The invention includes the ssRNAP itself, its fragments and fusion proteins, nucleic acids encoding it, specific promoters, and methods for its use in biotechnological applications such as in vitro RNA production, metagenomic screening, and mRNA therapeutic manufacturing.

Use CasesContent extracted from patent full text and abstract with AI.

• In vitro synthesis of mRNA for therapeutics, including vaccines and gene therapies.
• Cell-free protein expression systems for rapid screening and production of enzymes and proteins.
• Production of RNA/DNA hybrid molecules for diagnostic or research purposes.
• Large-scale manufacturing of mRNA vaccines and RNA drugs with improved process efficiency.
• Metagenomic screening and discovery of novel enzymes via cell-free systems.
• Expression of difficult-to-clone genes or toxic products outside living cells.
• Development of cell-free expression kits for synthetic biology, research, and diagnostics.
• High-throughput enzymatic screening and directed evolution workflows, including in droplets or microenvironments.

BenefitsContent extracted from patent full text and abstract with AI.

• Improved thermostability compared to T7 RNA polymerase, enabling effective transcription at higher temperatures and longer shelf-life.
• Faster transcription rates, resulting in shorter reaction times and greater productivity.
• Distinct promoter specificity that reduces unwanted cross-activity with common phage promoters, enabling orthogonal expression systems.
• High salt and temperature tolerance, useful for various reaction conditions and applications.
• Ability to incorporate non-canonical ribonucleotides (e.g., pseudouridine), which are critical for the synthesis of modified therapeutic mRNAs.
• Facilitates robust cell-free systems, broadening opportunities for enzyme discovery and functional screenings in metagenomics.
• Enables efficient production of RNA for diagnostics, therapeutics, and research, especially in demanding industrial settings.
• Supports the development of modular fusion proteins (e.g., with capping enzymes) for streamlined, one-step mRNA capping and transcription.

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Patent Abstract

The invention relates to a novel DNA-directed single-subunit RNA polymerase. The invention provides a single-subunit RNA polymerase, ssRNAP, the ssRNAP having advantageous features compared to the well-known T7 RNAP.