ArticlePloS one2022
Efficient multi-gene expression in cell-free droplet microreactors.
Article in PloS one, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Development of a Microdroplet-Based Functional Genomic Screening Pipeline by Combination of DNA Nanoflowers and PURExpress Cell-Free Expression.ACS synthetic biology · 2026Article
- A Programmable DNA Biohybrid Material Coupled with Intelligent Cell-Free Computation for Environmental Sensing.ACS omega · 2026Article
- Synthetic Biology of Plants and Microbes for Agriculture, Environment, and Future Applications.Chemical reviews · 2026Review
- A Guide to Biodetection in Droplets.Analytical chemistry · 2024Review
- Clonal Amplification-Enhanced Gene Expression in Synthetic Vesicles.ACS synthetic biology · 2023Article
- Cell-Free Production Systems in Droplet Microfluidics.Advances in biochemical engineering/biotechnology · 2023Review
- Solid-Phase Cell-Free Protein Synthesis and Its Applications in Biotechnology.Advances in biochemical engineering/biotechnology · 2023Article
Corrections and comments
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Authors and funding
3 authors.
Funding
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Abstract
Cell-free transcription and translation systems promise to accelerate and simplify the engineering of proteins, biological circuits and metabolic pathways. Their encapsulation on microfluidic platforms can generate millions of cell-free reactions in picoliter volume droplets. However, current methods struggle to create DNA diversity between droplets while also reaching sufficient protein expression levels. In particular, efficient multi-gene expression has remained elusive. We here demonstrate that co-encapsulation of DNA-coated beads with a defined cell-free system allows high protein expression while also supporting genetic diversity between individual droplets. We optimize DNA loading on commercially available microbeads through direct binding as well as through the sequential coupling of up to three genes via a solid-phase Golden Gate assembly or BxB1 integrase-based recombineering. Encapsulation with an off-the-shelf microfluidics device allows for single or multiple protein expression from a single DNA-coated bead per 14 pL droplet. We envision that this approach will help to scale up and parallelize the rapid prototyping of more complex biological systems.
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Registered trials
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