ArticleScientific reports2020
Rolling circle amplification of synthetic DNA accelerates biocatalytic determination of enzyme activity relative to conventional methods.
Article in Scientific reports, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 17 citations in OpenAlex.
- Transient Transfection of Rolling-Circle Amplified DNA in Biomanufacturing-Relevant Mammalian Cell Lines: A Comparison of Transfection Conditions for Optimal Protein Expression.Biotechnology and applied biochemistry · 2026Article
- Design-driven optimization of low-cost reagent formulations for reproducible and high-yielding cell-free gene expression.Nature communications · 2026Article
- Accessible biocatalyst development by rapidiScience · 2026Article
- Cell-Free Gene Expression: Methods and Applications.Chemical reviews · 2025Review
- Amplified DNA Heterogeneity Assessment with Oxford Nanopore Sequencing Applied to Cell Free Expression Templates.bioRxiv : the preprint server for biology · 2024Article
- Amplified DNA heterogeneity assessment with Oxford Nanopore sequencing applied to cell free expression templates.PloS one · 2024Article
- HyperXpress: Rapid Single Vessel DNA Assembly and Protein Production in Microliterscale.Frontiers in bioengineering and biotechnology · 2022Article
- Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems.Journal of visualized experiments : JoVE · 2021Article
- Study on Factors Affecting the Performance of a CRISPR/Cas-Assisted New Immunoassay: Detection of Salivary Insulin as an Example.Frontiers in bioengineering and biotechnology · 2021Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The ability to quickly and easily assess the activity of large collections of enzymes for a desired substrate holds great promise in the field of biocatalysis. Cell-free synthesis, although not practically amenable for large-scale enzyme production, provides a way to accelerate the timeline for screening enzyme candidates using small-scale reactions. However, because cell-free enzyme synthesis requires a considerable amount of template DNA, the preparation of high-quality DNA "parts" in large quantities represents a costly and rate-limiting prerequisite for high throughput screening. Based on time-cost analysis and comparative activity data, a cell-free workflow using synthetic DNA minicircles and rolling circle amplification enables comparable biocatalytic activity to cell-based workflows in almost half the time. We demonstrate this capability using a panel of sequences from the carbon-nitrogen hydrolase superfamily that represent possible green catalysts for synthesizing small molecules with less waste compared to traditional industrial chemistry. This method provides a new alternative to more cumbersome plasmid- or PCR-based protein expression workflows and should be amenable to automation for accelerating enzyme screening in industrial applications.
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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.