ArticleNucleic acids research2023
High-salt transcription from enzymatically gapped promoters nets higher yields and purity of transcribed RNAs.
Article in Nucleic acids research, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 25 citations in OpenAlex.
- IntegratedMolecular therapy. Nucleic acids · 2026Article
- The current landscape of mRNA therapy and the strategies for mRNA purification and dsRNA removal.Journal of biomedical science · 2026Review
- Active-learning-guided optimization of cell-free systems for genome-wide transcriptomic profiling reveals progressive layers of regulation.Nature communications · 2026Article
- DNA Flap-Mediated Control of Transcription for Programmable RNA Synthesis.Angewandte Chemie (International ed. in English) · 2026Article
- Directed Evolution of T7 RNA Polymerase Minimizes dsRNA By-product and Enables High-Fidelity mRNA Synthesis for Demanding Therapeutic Applications.Research (Washington, D.C.) · 2026Article
- Impact of RNA extraction on respiratory microbiome analysis using third-generation sequencing.BMC genomics · 2025Article
- Continuous in situ synthesis of a complete set of tRNAs sustains steady-state translation in a recombinant cell-free system.Nature communications · 2025Article
- Removal of dsRNA byproducts using affinity chromatography.Molecular therapy. Nucleic acids · 2025Article
- A new approach to RNA synthesis: immobilization of stably and functionally co-tethered promoter DNA and T7 RNA polymerase.Nucleic acids research · 2024Article
- DNA-terminus-dependent transcription by T7 RNA polymerase and its C-helix mutants.Nucleic acids research · 2024Article
- Rational design of oligonucleotides for enhanced in vitro transcription of small RNA.RNA (New York, N.Y.) · 2024Article
- Leveraging Active Learning to Establish Efficient In Vitro Transcription and Translation from Bacterial Chromosomal DNA.ACS omega · 2024Article
- Bacteriophage RNA polymerases: catalysts for mRNA vaccines and therapeutics.Frontiers in molecular biosciences · 2024Review
- Article
- Understanding the impact ofFrontiers in molecular biosciences · 2024Review
- A simple approach to improving RNA synthesis: Salt inhibition of RNA rebinding coupled with strengthening promoter binding by a targeted gap in the DNA.Methods in enzymology · 2023Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
T7 RNA polymerase is commonly used to synthesize large quantities of RNA for a wide variety of applications, from basic science to mRNA therapeutics. This in vitro system, while showing high fidelity in many ways, is also well known for producing longer than encoded RNA products, particularly under high-yield reaction conditions. Specifically, the resulting product pool is contaminated by an often disperse collection of longer cis-primed extension products. In addition to reducing yield via the conversion of correctly encoded RNA to longer products, self-primed extension generates partially double-stranded RNAs that can trigger the innate immune response. Extensive and low-yield purifications are then required to produce therapeutic RNA. Under high-yield conditions, accumulating concentrations of RNA effectively compete with promoter DNA for polymerase binding, driving self-primed extension at the expense of correct initiation. In the current work, we introduce a simple and novel modification in the DNA to strengthen promoter binding, shifting the balance back toward promoter-driven synthesis and so dramatically reducing self-primed extension. The result is higher yield of the encoded RNA at the outset and reduced need for extensive purifications. The approach can readily be applied to the synthesis of mRNA-length products under high-yield conditions.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.