We demonstrate utility of AI-2 Controllers by modulating naturally occurring processes of biofilm formation. We envision that 'controller cells' that modulate bacterial behavior by manipulating molecular communication, will find use in a variety of applications, particularly those employing natural or synthetic bacterial consortia.
This summer, our team successfully constructed two cell machines to manipulate the molecular connection between cells by modulating the bacterial 'universal language', autoinducer-2: AI-2 Supplier is the cell machine which can directly supply and enrich the AI-2 molecular level; AI-2 Consumer is another cell machine which can sense, absorb and degrade the AI-2 in the environment.
We demonstrate utility of AI-2 Controllers by modulating naturally occurring processes of biofilm formation. We envision that "controller cells" that modulate bacterial behavior by manipulating molecular communication, will find use in a variety of applications, particularly those employing natural or synthetic bacterial consortia.
Our goals
AI-2 Supplier
To enrich the AI-2 concentration in the nature or artificial environment, we constructed two AI-2 Supplier Devices by overexpression of the components responsible for AI-2 production (luxS, mtn).
AI-2 Consumer
To 'quench' AI-2 signal in the nature or artificial environment, we constructed six AI-2 Consumer Devices by overexpression the components responsible for AI-2 uptake(lsrACDB), phosphorylation(lsrK) and degradation (lsrFG).
AI-2 Response Device
We successfully constructed two devices that could respond to AI-2 by producing GFP, to provide an independent means to use AI-2 Controller to alter heterologous gene expression. we show that a 1:1 mixture of AI-2 Response Device with AI-2 suppliers activated QS-activated GFP expression from the control group. And 1:1 mixture of AI-2 Response Device with AI-2 consumers could significantly depress QS-activated gene expression from the control cells.
Biosafety Considerations
This summer, we plan to mf-lon ssrA tag into 5 essential genes by using CRIPSR/Cas9 technology. By applying gene circuits to control mf-lon protease expression under the assigned biocontainment conditions, we can blocks essential gene expression to kill the cell upon loss of the biocontainment signal.
Demonstration of Experiment Results
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