Supplementary Materials

Supplementary Material for:

Detection of pathological biomarkers in human clinical samples via amplifying genetic switches and logic gates

Alexis Courbet, Drew Endy, Eric Renard, Franck Molina,* Jérôme Bonnet*

*Corresponding author. E-mail: jerome.bonnet{at}inserm.fr (J.B.); franck.molina{at}sys2diag.cnrs.fr (F.M.)

Published 27 May 2015, Sci. Transl. Med. 7, 289ra83 (2015)
DOI: 10.1126/scitranslmed.aaa3601

This PDF file includes:

  • Materials and Methods
  • Notes
  • Fig. S1. Conceptual workflow for the systematic development of medical bactosensors.
  • Fig. S2. Bacterial chassis growth characteristics in urine and serum.
  • Fig. S3. Bacterial chassis viability in urine and serum from single-cell measurements.
  • Fig. S4. Influence of clinical media (urine and serum) on GFP fluorescence output generation and measurement.
  • Fig. S5. Population and single-cell measurements of multiplexing Boolean integrase logic gates operation in urine and serum.
  • Fig. S6. Stability of genetic memory in clinical samples by addressing DNA register with Sanger sequencing.
  • Fig. S7. Workflow for engineering switches that respond to biological signals over user-defined thresholds.
  • Fig. S8. Kinetic measurements and transfer functions of promoters and switches.
  • Fig. S9. Multiplexing detection of glucose and NOx, with AND-BCD-RFP, NAND-BCD-RFP, and NOR-BCD-RFP Boolean integrase logic gates.
  • Fig. S10. Single-cell measurements of fold changes and DERs for promoters and switches.
  • Fig. S11. Evaluation of the robustness of inducible systems against clinical media-induced perturbation (urine and serum).
  • Fig. S12. Comparison of the operational characteristic of AND logic gates with and without the bicistronic device (BCD) for their use in clinical samples.
  • Fig. S13. Insulation of bactosensors in hydrogel beads and pathological signal detection.
  • Fig. S14. ROC analysis for bactosensor-mediated detection of glucose in urine.
  • Fig. S15. Potential modalities of bactosensor-based diagnosis and composability of integrase-based logic for the development of decision-making tests.
  • Data file S1. Numerical values of OD600 and fluorescence from plate reader measurements and calculated RPUs used to develop plots and figures.
  • Note S1. Experimental design: Use of reference promoters.
  • Note S2. Genetic design: Use of standardized genetic elements.
  • Note S3. Bacterial encapsulation toward a clinical format.
  • References (6571)

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