ArticleChembiochem : a European journal of chemical biology2019
Evolution of C-Terminal Modification Tolerance in Full-Length and Split T7 RNA Polymerase Biosensors.
Article in Chembiochem : a European journal of chemical biology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed.
- Engineered chimeric T7 RNA polymerase improves salt tolerance and reduces dsRNA impurity generation during in vitro transcription of mRNA.Nucleic acids research · 2025Article
- A modular cell-free protein biosensor platform using split T7 RNA polymerase.Science advances · 2025Article
- Current biosensing strategies based on in vitro T7 RNA polymerase reaction.Biotechnology notes (Amsterdam, Netherlands) · 2025Review
- A modular cell-free protein biosensor platform using split T7 RNA polymerase.bioRxiv : the preprint server for biology · 2024Article
- Continuous directed evolution of a compact CjCas9 variant with broad PAM compatibility.Nature chemical biology · 2024Article
- Virus-assisted directed evolution of biomolecules.Current opinion in chemical biology · 2023Review
- Engineering Rapalog-Inducible Genetic Switches Based on Split-T7 Polymerase to Regulate Oncolytic Virus-Driven Production of Tumour-Localized IL-12 for Anti-Cancer Immunotherapy.Pharmaceuticals (Basel, Switzerland) · 2023Article
- Phage-Assisted Continuous Evolution and Selection of Enzymes for Chemical Synthesis.ACS central science · 2021Article
- A System for the Evolution of Protein-Protein Interaction Inducers.ACS synthetic biology · 2021Article
- Contingency and chance erase necessity in the experimental evolution of ancestral proteins.eLife · 2021Article
- Phage-assisted continuous and non-continuous evolution.Nature protocols · 2020Article
- Continuous evolution of SpCas9 variants compatible with non-G PAMs.Nature biotechnology · 2020Article
- Split T7 RNA polymerase biosensors to study multiprotein interaction dynamics.Methods in enzymology · 2020Article
- Development of a Split Esterase for Protein-Protein Interaction-Dependent Small-Molecule Activation.ACS central science · 2019Article
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Authors and funding
3 authors.
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Abstract
T7 RNA polymerase (RNAP) is a powerful protein scaffold for the construction of synthetic biology tools and biosensors. However, both T7 RNAP and its split variants are intolerant to C-terminal modifications or fusions, thus placing a key limitation on their engineering and deployment. Here, we use rapid continuous-evolution approaches to evolve both full-length and split T7 RNAP variants that tolerate modified C termini and fusions to entire other proteins. Moreover, we show that the evolved split C-terminal RNAP variants can function as small-molecule biosensors, even in the context of large C-terminal fusions. This work provides a panel of modified RNAP variants with robust activity and tolerance to C-terminal fusions, and provides insights into the biophysical requirements of the C-terminal carboxylic acid functional group of T7 RNAP.
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