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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.

Updated 4 cited papers83 editable layers

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At a glance

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 (16:9), 83 editable layers, 2 generated objects

Step by step

  1. DNA inA plasmid or linear DNA template encoding the protein of interest goes into the reaction (Gregorio et al. 2019)
  2. Open reactionA 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 outProtein 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. UsesFreeze-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

Papers this template follows

  1. Shimizu et al., “Cell-free translation reconstituted with purified components”, Nat. Biotechnol. 2001. doi:10.1038/90802method origin
  2. Gregorio, Levine & Oza, “A User's Guide to Cell-Free Protein Synthesis”, Methods Protoc. 2019. doi:10.3390/mps2010024review
  3. Pardee et al., “Paper-based synthetic gene networks”, Cell 2014. doi:10.1016/j.cell.2014.10.004primary
  4. Pardee et al., “Portable, on-demand biomolecular manufacturing”, Cell 2016. doi:10.1016/j.cell.2016.09.013primary

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).

How templates are written and checked: editorial guidelines.

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.

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cell-free protein synthesisCFPSin vitro transcription translationPURE systemcell extractTX-TLribosomeT7 RNA polymeraseopen systemfreeze-driedbiosensoron-demand manufacturingrapid prototypinggraphical abstract3D model

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