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Establishing Baseline Performance Scores for the CAGEN Robust Gene Response Challenge

This study aims to establish baseline performance scores for the Cagen Robust Gene Response Challenge, evaluating the robustness and speed of genetically engineered networks in inducing gene responses. Participants will assess selected designs based on metrics like protein levels and time integral of square error. The challenge involves inducing YFP expression in response to IPTG with low variability across cells and temperatures. Future work includes refining single-cell measurements using microfluidics.

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Establishing Baseline Performance Scores for the CAGEN Robust Gene Response Challenge

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  1. Establishing Baseline Performance Scores for the CAGEN Robust Gene Response Challenge Shaunak Sen Caltech CMS Oct 11, 2011

  2. CAGEN - Critical Assessment of • Genetically Engineered Networks • Systematize the design of genetic networks • (robustness, connectability) • - Facilitate applications, for ex. in agriculture, medicine

  3. CAGEN Challenge #1 : Robust Gene Response Induce OUTPUT (YFP) in response to INPUT (IPTG), so that, Detectable: >10 fold induction Robust: Low variability across cells & temperatures Fast: Quick response Speed >10 X0 Low Variability OUTPUT YFP induction 1 X0 Time IPTG Addition INPUT

  4. PurposeEstablish Baseline Performance Challenge Specifications Purpose: Establish Baseline Performance ANNOUNCEMENT Self Assessment by Participants SELECTION Critical Assessment of Selected Designs

  5. Performance metric to score robustness and speed of designs r(t) yj(t) j=2…15 Protein Levels (nM) simulation Time integral of square error Worst Case Time (hours) X Px x Normalizing factor (equilibrium amplitude of reference trace)

  6. Single cell dynamics are measured for a simple reference design, for different temperatures #2 #1 Pinducible YFP E. coli Plac Plac cI-yfp yfp IPTG IPTG

  7. Movies Pinducible YFP

  8. Reference trace exhibits > 10x induction Cell division Mean Fluorescence (a.u.) 10x induction No induction Camera Bgd. Time (minutes)

  9. Performance scores for inducible protein expression circuits #2 #1 Reference trace (32 °C) Traces at 32 °C Traces at 34 °C Traces at 30 °C Reference trace (34 °C) Traces at 34 °C Traces at 30 °C S = 0.27 S = 0.29 Mean Fluorescence (a.u.) Mean Fluorescence (a.u.) Plac Plac cI-yfp yfp IPTG IPTG Time (minutes) Time (minutes)

  10. Population level metric to estimate performance score r(t) Single-Cell Dynamics/ Microscopy yj(t) j=2…15 Population Dynamics/ Flow Cytometry Protein Levels (nM) Count simulation Time (minutes) Protein Levels (nM) Time (hours)

  11. Future WorkRefined single-cell measurements using μfluidics Challenge Specifications Simulation/ Eric Klavins Baseline for Critical Assessment Self Assessment by Participants Critical Assessment of Selected Designs Benefit #2: Similar decay time measurement. Benefit #1. Exact induction time measurement.

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