Synchronous long-term oscillations in a synthetic gene circuit · 2016. 11. 14. · Title:...

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.