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Synchronous long-term oscillations in a syntheticgene circuitEMILEE HOLTZAPPLE
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Outline•Background
•Methods
•Results
•Future Directions
•Conclusions
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BackgroundFirst described in “A synthetic oscillatory network of transcriptional regulators” (Elowitz and Leibler 2000)
• Relationship between repressor concentration and transcription rate
• Translation rate• Protein and mRNA decay
rate
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Stochastic
Gillespie’s Direct
Deterministic
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Methods
For the first experiment, the original plasmid designs were used
Low-copy pSC101 plasmid and high-copy ColE1 reporter
Microfluidic device was used to control cell population
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Production rate
Biased autocorrelation function
Unbiased autocorrelation function
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Power spectrum analysis
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Results
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ConclusionsOriginal variation in the repressilator can be partially attributed to insufficient technology and extraneous elements
Removal of high-copy cloning vectors, degradation tags, and addition of a
A simple circuit can produce regular oscillations, even when subjected to intrinsic cellular noise
Study synthetic circuits with both positive and negative elements
Apply methodology to other synthetic circuits to improve performance
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Original repressilator:
Elowitz, M. B. and S. Leibler (2000). "A synthetic oscillatory network of transcriptional regulators." Nature 403(6767): 335-338.