Team:Paris Bettencourt/Project/Phage Sensor

Overview

We develop a sensor to target antibiotic resistances in tuberculosis to test if a specific strain carries a certain antibiotic resistance. Our sensor system consists out of a phagemid with a CRISPR/Cas system and LacZ as a reporter under the control of a pREC promoter (the promoter of the RecA protein that is involved in the stress response of bacteria). If our CRISPR/Cas system can now bind to the target (antibiotic resistance gene), the Cas9 generates at this specific target site a double strand break, which then starts the expression of our reporter, as the promoter gets active at stress that results from double strand breaks. Because our system is on a phagemid, the sensor system will be spread all over the population, which will give a clear color output if the target has been detected. This means, depending on what target sequence our system carries, we can identify the different antibiotic resistances that a strain might carry. This is a novel way of detecting resistances in bacterial strains. As a proof of concept we will use E.coli to target KanR. The used phage will be M13. This sensor could potentially be used to test if a patient has TB and what type of resistance genes the specific strain contains to adapt drug treatment.