Experiments and Results

With the overarrching goal of prooducing a modular and scalable electrogenetic toolkit, we developped genetic construcs and custom hardware and software.

Experiments fall in the following categories: Genetic engieering where we designed and characterised genetic construcst.
A serie of experiments aiming at screening a library of E. coli promoter for Electro-Genetic sensitivity.
A serie of experiemtn to characterise our hardware and software

Experiments fall in the following categories:

  • Genetic engineering where we designed and characterised genetic construcst.
  • A serie of experiments aiming at screening a library of E. coli promoter for Electro-Genetic sensitivity.
  • A serie of experiemtn to characterise our hardware and software

Main Acheivements

  1. We delivered a characterised part collection enabling Electro-genetics in iGEM.
  2. We successfully screened a library of promoter in E. coli for Electrically inducible Gene expression and added 19 responsive promoters to our collection.
  3. We developed specific and new Hardware enabling Electro-genetics, at low cost, in iGEM and in context beyond synthetic biology.
  4. We described and characterised genetic circuits performing electrical signal processing function in E. coli.



Genetic engineering

Here we seek to develop and characterise the 3-plasmid system part by part and as a system

Input characterisation

The input is designed to receive a signal that is then transmitted to the processor. First, we constructed the input to receive an electrical signal, and, in turn, activate the transcriptional unit coding for the processor transcription factor (CinR); however, upon reviewing the literature, we later discovered a different electrical inducible promotor known as pSoxS, a piocynin inducible promotor that have already been used in iGEM, which we deciede to implement instead.

Processor characterisation

The processor is the part in our device responsible for transmitting the signal from the input to the output. Any signal passing through the processor can be modulated, allowing for the same input to have multiple different outputs. Our processor is comprised two main parts: a (1) promotor (Pcin), which responds to the signal of the input and a (2) chemical inducer (OHC14) with a transcription unit created to communicate the signal to the output via activation of its promotor (pLUX) with a transcriptional factor(LuxRQ).

Output characterisation

The output is designed to transform the message from the processor into a signal readable to our measurement tools. One output construct we designed is comprised of a promotor (pLuxR) sensitive to the processor’s message, and a transcriptional unit expressing an RFP or a Lysis gene. (The lysis gene destroys the cells, releasing a sufficient concentration of ions to read a signal on our potentiostat.)

Multiple plasmid characterisation

The multi-plasmid device is made of 3 plasmids that each code for either the input, the processor and the output. This characterisation was designed and purposed to prove that the 3 plasmids can communicate with each other and that the signal they pass to each other can be modulated with chemicals products.



Electro-Resonsive Promtoters screening

Hardware

We screened 500 promoters in E. coli for there response to exposure to electrical signals.
We found 19 responsive promoters (8 induced and 11 repressed).
To the part collection

To interface population of engineered bacterial with electronic systems as well as to prototype and run experiement, we delivered a working Hardware Collection

Electro Library Screening

Hardware Experiments
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