ArticleG3 (Bethesda, Md.)2022
Engineering efficient termination of bacteriophage T7 RNA polymerase transcription.
Article in G3 (Bethesda, Md.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 36 citations in OpenAlex.
- Exploring the Evolutionary Landscape with Targeted In Vivo Hypermutations.Biomedicines · 2026Review
- A self-replicating artificial module-genome that generates bacterial chromosome replication system in vitro.Nucleic acids research · 2026Article
- Enhanced dsRNA Production via a Three-Terminator Vector and Transcriptomic Correlates of RNAi Exposure in Thrips.Insects · 2026Article
- Identification of Phage RNA Polymerases That Minimize Double-Stranded RNA By-Product Formation and Their Characterization via In Vitro Transcription.Microorganisms · 2026Article
- Expression of nano-engineered RNA organelles in bacteria.Nature communications · 2026Article
- Process and analytical strategies for the safe production of mRNA vaccines and therapeutics.Molecular biology reports · 2026Review
- Biomimetic peptide conjugates as emerging strategies for controlled release from protein-based materials.Drug delivery · 2025Review
- Protocol for inducible expression of small non-coding RNAs using a tet-on system in Saccharomyces cerevisiae.STAR protocols · 2025Article
- Comparative analysis of convergent and divergent T7 RNA polymerase promoters for the synthesis of dsRNA in vivo and in vitro.RNA (New York, N.Y.) · 2025Article
- Simultaneous in vitro expression of minimal 21 transfer RNAs by tRNA array method.Nature communications · 2025Article
- Continuous in situ synthesis of a complete set of tRNAs sustains steady-state translation in a recombinant cell-free system.Nature communications · 2025Article
- Design and application of expression constructs for FMDV serotype O structural proteins.Biotechnology letters · 2025Article
- Free introns of tRNAs as complementarity-dependent regulators of gene expression.Molecular cell · 2025Article
- Regulatory Components for Bacterial Cell-Free Systems Engineering.ACS synthetic biology · 2024Review
- Discrete measurements of RNA polymerase and reverse transcriptase fidelity reveal evolutionary tuning.RNA (New York, N.Y.) · 2024Article
- Characterization of the dual regulation by a c-di-GMP riboswitch Bc1 with a long expression platform fromMicrobiology spectrum · 2024Article
- Effective Synthesis of High-Integrity mRNA Using In Vitro Transcription.Molecules (Basel, Switzerland) · 2024Article
- Development of advanced control material for reverse transcription-mediated bacterial nucleic acid amplification tests.Journal of clinical microbiology · 2024Article
- Single-molecule RNA sizing enables quantitative analysis of alternative transcription termination.Nature communications · 2024Article
- Rapid Antibacterial Activity Assessment of Chimeric Lysins.International journal of molecular sciences · 2024Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
The bacteriophage T7 expression system is one of the most prominent transcription systems used in biotechnology and molecular-level research. However, T7 RNA polymerase is prone to read-through transcription due to its high processivity. As a consequence, enforcing efficient transcriptional termination is difficult. The termination hairpin found natively in the T7 genome is adapted to be inefficient, exhibiting 62% termination efficiency in vivo and even lower efficiency in vitro. In this study, we engineered a series of sequences that outperform the efficiency of the native terminator hairpin. By embedding a previously discovered 8-nucleotide T7 polymerase pause sequence within a synthetic hairpin sequence, we observed in vivo termination efficiency of 91%; by joining 2 short sequences into a tandem 2-hairpin structure, termination efficiency was increased to 98% in vivo and 91% in vitro. This study also tests the ability of these engineered sequences to terminate transcription of the Escherichia coli RNA polymerase. Two out of 3 of the most successful T7 polymerase terminators also facilitated termination of the bacterial polymerase with around 99% efficiency.
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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.