# Cell-free protein synthesis: DNA in, protein out

> A pipeline graphical abstract of cell-free protein synthesis. A DNA template goes into an open reaction — a cell extract or the reconstituted PURE system — which transcribes and translates it in one tube, with no cell wall or genome in the way, and a protein comes out within hours. Freeze-dried, the same reaction becomes portable for rapid prototyping, paper biosensors and on-demand production.

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- Open in the LabCanvas editor (LabCanvas account): https://www.labcanvas.com/editor.html?template=cell-free-protein-synthesis
- Updated: 2026-10-14
- Category: Graphical abstracts
- Method: Graphical abstract
- Starting materials: A DNA template (plasmid or linear) plus a cell extract or the PURE system
- Product: A finished protein in hours, in an open tube — no living cell
- Key result: ~160 µg/ml/h in PURE batch (Shimizu et al. 2001); E. coli reactions scale 10 µL → 100 L, in-house extract ~$0.019/µL, freeze-dried extract keeps for months at room temperature (Gregorio et al. 2019)
- Figure: 1920 × 1080 px, 83 editable layers (2 generated objects with settings)

## Steps

1. **DNA in**: A plasmid or linear DNA template encoding the protein of interest goes into the reaction (Gregorio et al. 2019)
2. **Open reaction**: A cell extract or the reconstituted PURE system (T7 RNA polymerase, 20 aminoacyl-tRNA synthetases, ribosomes, tRNAs, amino acids, energy) transcribes and translates it; no membrane or genome, so all the energy goes to the protein (Shimizu et al. 2001; Gregorio et al. 2019)
3. **Protein out**: Protein appears within hours — the PURE system makes ~160 µg/ml/h — and tagged factors are removed by affinity chromatography in ~1 h (Shimizu et al. 2001)
4. **Uses**: Freeze-dried and rehydrated, the reaction is portable: rapid prototyping of genetic circuits, paper biosensors with colour readouts, and on-demand therapeutics (Pardee et al. 2014, 2016)

## Uses

- Graphical abstracts on cell-free synthetic biology
- Talks on PURE vs extract-based CFPS
- Teaching in vitro transcription–translation and on-demand biomanufacturing

## References

1. Shimizu et al., “Cell-free translation reconstituted with purified components”, Nat. Biotechnol. 2001. https://doi.org/10.1038/90802 (method origin)
2. Gregorio, Levine & Oza, “A User's Guide to Cell-Free Protein Synthesis”, Methods Protoc. 2019. https://doi.org/10.3390/mps2010024 (review)
3. Pardee et al., “Paper-based synthetic gene networks”, Cell 2014. https://doi.org/10.1016/j.cell.2014.10.004 (primary)
4. Pardee et al., “Portable, on-demand biomolecular manufacturing”, Cell 2016. https://doi.org/10.1016/j.cell.2016.09.013 (primary)

Credit: Drawn by LabCanvas after Shimizu et al., Nat. Biotechnol. 2001 (doi:10.1038/90802); Gregorio, Levine & Oza, Methods Protoc. 2019 (doi:10.3390/mps2010024); and Pardee et al., Cell 2014 (doi:10.1016/j.cell.2014.10.004) and 2016 (doi:10.1016/j.cell.2016.09.013).

## Questions

**What does the “Cell-free protein synthesis: DNA in, protein out” template show?** A pipeline graphical abstract of cell-free protein synthesis. A DNA template goes into an open reaction — a cell extract or the reconstituted PURE system — which transcribes and translates it in one tube, with no cell wall or genome in the way, and a protein comes out within hours. Freeze-dried, the same reaction becomes portable for rapid prototyping, paper biosensors and on-demand production.

**What are the steps shown in “Cell-free protein synthesis: DNA in, protein out”?** 1. DNA in: A plasmid or linear DNA template encoding the protein of interest goes into the reaction (Gregorio et al. 2019). 2. Open reaction: A cell extract or the reconstituted PURE system (T7 RNA polymerase, 20 aminoacyl-tRNA synthetases, ribosomes, tRNAs, amino acids, energy) transcribes and translates it; no membrane or genome, so all the energy goes to the protein (Shimizu et al. 2001; Gregorio et al. 2019). 3. Protein out: Protein appears within hours — the PURE system makes ~160 µg/ml/h — and tagged factors are removed by affinity chromatography in ~1 h (Shimizu et al. 2001). 4. Uses: Freeze-dried and rehydrated, the reaction is portable: rapid prototyping of genetic circuits, paper biosensors with colour readouts, and on-demand therapeutics (Pardee et al. 2014, 2016).

**What are the starting materials and the product?** Starting materials: A DNA template (plasmid or linear) plus a cell extract or the PURE system. Product: A finished protein in hours, in an open tube — no living cell.

**Which papers is “Cell-free protein synthesis: DNA in, protein out” based on?** Shimizu et al., “Cell-free translation reconstituted with purified components”, Nat. Biotechnol. 2001 (doi:10.1038/90802); Gregorio, Levine & Oza, “A User's Guide to Cell-Free Protein Synthesis”, Methods Protoc. 2019 (doi:10.3390/mps2010024); Pardee et al., “Paper-based synthetic gene networks”, Cell 2014 (doi:10.1016/j.cell.2014.10.004); Pardee et al., “Portable, on-demand biomolecular manufacturing”, Cell 2016 (doi:10.1016/j.cell.2016.09.013).

**Can I edit the “Cell-free protein synthesis: DNA in, protein out” template?** Yes. Open it in the LabCanvas editor: all 83 layers are editable, including 2 generated objects (structures, plots, equipment) with their own settings. Change the text, colours and layout, then export PNG, TIFF, SVG or PDF.

**Do I need an account to use this template?** Not to view it here. To open and edit it, sign in to LabCanvas or create a free account (the free plan keeps up to 3 designs). Downloading the figure is part of LabCanvas Pro (US$19 a month or US$199 a year).