synchronous long-term oscillations in a synthetic gene circuit · 2016. 11. 14. · title:...
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Synchronous long-term oscillations in a syntheticgene circuitEMILEE HOLTZAPPLE
Outline•Background
•Methods
•Results
•Future Directions
•Conclusions
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
Stochastic
Gillespie’s Direct
Deterministic
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
Production rate
Biased autocorrelation function
Unbiased autocorrelation function
Power spectrum analysis
Results
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
Original repressilator:
Elowitz, M. B. and S. Leibler (2000). "A synthetic oscillatory network of transcriptional regulators." Nature 403(6767): 335-338.