The Detector Synthetic Cell detects the presence of two chemicals and produces an intracellular fluorescent signal in response. A two-input detector cell demonstrates the ability to detect multiple small molecules, balance expression dynamics, and implement simple genetic logic within a liposome.

Implementing these modules requires synthetic cells capable of small molecule transport across the membrane. To this end, we have integrated the alpha-hemolysin membrane protein (Alpha hemolysin) into our liposomes (“Hello, world” PURE Liposomes)

IPTG detector

This cell detects IPTG in the environment and produces a fluorescent signal in response.

Uses these modules:

Detector Cell_ lacI_white.png

Tetracycline detector

This cell detects aTc in the environment and produces a fluorescent signal in response.

Uses these modules:

Detector Cell_ tetR_white.png

IPTG-aTc (AND gate) detector

The IPTG-aTc detector integrates both Lac and Tet repression. This allows for control of expression using both inducer molecules, with maximum expression occurring when both inducers are present—a logical AND gate.

Uses these modules:

  1. IPTG detector (LacI Inducible Module)
  2. Tetracycline detector (TetR Inducible Module)
  3. IPTG-aTc (AND gate) detector (Lac-Tet inducible AND gate expression)

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<aside> <img src="/icons/wrench_gray.svg" alt="/icons/wrench_gray.svg" width="40px" /> Usage

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Protocols and parameters for building and using the detector cells are coming soon.

<aside> <img src="/icons/checklist_gray.svg" alt="/icons/checklist_gray.svg" width="40px" /> Performance Data

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Alpha-hemolysin permeabilization of the liposome membrane

30min_timelapse_cropped.mp4

Liposomes containing a green pyranine dye were permeabilized by adding purified alpha-hemolysin to the outer solution. The dye remains contained within the liposomes until alpha-hemolysin inserts; the dye then leaks into the outer solution.

TetR Detector Cell

tetR detector synthetic cell induced at multiple anhydrotetracyline concentrations. 8 timepoints are displayed per sample condition, with approximately 22 minutes per timepoint. The detector cells were incubated for a total of 12 hours. First row: induction of tetR detectors using 625 nM, 312.5 nM, and 0 nM (fully repressed) anhydrotetracycline, introduced into the outer buffer. Second row: induction of tetR detector with 2500 nM anhydrocycline in the inner solution, and positive control without tetR repression.

tetR detector synthetic cell induced at multiple anhydrotetracyline concentrations. 8 timepoints are displayed per sample condition, with approximately 22 minutes per timepoint. The detector cells were incubated for a total of 12 hours. First row: induction of tetR detectors using 625 nM, 312.5 nM, and 0 nM (fully repressed) anhydrotetracycline, introduced into the outer buffer. Second row: induction of tetR detector with 2500 nM anhydrocycline in the inner solution, and positive control without tetR repression.

GFP expression within synthetic cells when induced with 312.5 uM anhydrotetracycline.

GFP expression within synthetic cells when induced with 312.5 uM anhydrotetracycline.

TetR detector cells induced with 325 nM anhydrotetracycline.

TetR detector cells induced with 325 nM anhydrotetracycline.