ArticleBiotechnology and bioengineering2017
Short DNA containing χ sites enhances DNA stability and gene expression in E. coli cell-free transcription-translation systems.
Article in Biotechnology and bioengineering, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers, 1 of them a synthesis that pooled it.
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Who cites it
44 citing papers in PubMed, 1 synthesis or guideline pooled it, 113 citations in OpenAlex.
- Cell-Free Approach for Non-canonical Amino Acids Incorporation Into Polypeptides.Frontiers in bioengineering and biotechnology · 2020Pooled it
- One-pot cloning and protein expression platform for genetic engineering.bioRxiv : the preprint server for biology · 2025Article
- Regulating Protein Immobilization During Cell-Free Protein Synthesis in Hyaluronan Microgels.Advanced biology · 2025Article
- Cell-free expression system: a promising platform for bacteriophage production and engineering.Microbial cell factories · 2025Review
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Modular Golden Gate Assembly of Linear DNA Templates for Cell-Free Prototyping.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Purification and in vivo, cell-free, and in vitro characterization of CRISPR-Cas12a2.Methods in enzymology · 2025Article
- Clonal Amplification-Enhanced Gene Expression in Synthetic Vesicles.ACS synthetic biology · 2023Article
- Established and Emerging Methods for Protecting Linear DNA in Cell-Free Expression Systems.Methods and protocols · 2023Review
- Short Activators and Repressors of RNA Toehold Switches.ACS synthetic biology · 2023Article
- T7Max transcription system.Journal of biological engineering · 2023Article
- The Use of Cell-free Protein Synthesis to Push the Boundaries of Synthetic Biology.Biotechnology and bioprocess engineering : BBE · 2023Review
- Cell-Free Display Techniques for Protein Evolution.Advances in biochemical engineering/biotechnology · 2023Article
- Solid-Phase Cell-Free Protein Synthesis and Its Applications in Biotechnology.Advances in biochemical engineering/biotechnology · 2023Article
- Constructing Cell-Free Expression Systems for Low-Cost Access.ACS synthetic biology · 2022Article
- A TXTL-Based Assay to Rapidly Identify PAMs for CRISPR-Cas Systems with Multi-Protein Effector Complexes.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Article
- Best Practices for DNA Template Preparation Toward Improved Reproducibility in Cell-Free Protein Production.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Protocell arrays for simultaneous detection of diverse analytes.Nature communications · 2021Article
- NgAgo possesses guided DNA nicking activity.Nucleic acids research · 2021Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Escherichia coli cell-free transcription-translation (TXTL) systems offer versatile platforms for advanced biomanufacturing and for prototyping synthetic biological parts and devices. Production and testing could be accelerated with the use of linear DNA, which can be rapidly and cheaply synthesized. However, linear DNA is efficiently degraded in TXTL preparations from E. coli. Here, we show that double-stranded DNA encoding χ sites-eight base-pair sequences preferentially bound by the RecBCD recombination machinery-stabilizes linear DNA and greatly enhances the TXTL-based expression and activity of a fluorescent reporter gene, simple regulatory cascades, and T7 bacteriophage particles. The χ-site DNA and the DNA-binding λ protein Gam yielded similar enhancements, and DNA with as few as four χ sites was sufficient to ensure robust gene expression in TXTL. Given the affordability and scalability of producing the short χ-site DNA, this generalized strategy is expected to advance the broad use of TXTL systems across its many applications. Biotechnol. Bioeng. 2017;114: 2137-2141. © 2017 Wiley Periodicals, Inc.
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