Integrated Human Practices(Gold)

Integrated Human Practices

Prototype

The device prototype is designed in such a way that it ensures the physical containment of the construct. It is similar to a 50mL falcon tube with a one way lock which ensures that the device cannot be opened once it is capped tightly. This device will contain the phasmids that can infect Mycobacterium and dispense the gene circuits into them along with middle brook media containing IPTG and Arabinose, all of which is separated from the exterior by an insoluble membrane. After adding the pathological sample into the device, a small tube will be added to the device to pierce the membrane on closing the cap. This ensures the mixing of the media in the device with the pathological sample.

Biosafety Module

One of the major concerns raised by many of our audience was the accidental chances of the device getting released into the wild. In-order to ensure the biosafety of the module in cases of such accidents, we are planning to incorporate a biosafety module in our device. The biosafety module involves a Nuclease, which chews up the DNA and RNA of the host when activated. This nuclease will be under the control of a highly repressed promoter, repressed by a protein R. This protein R will be under the control of an inducible promoter. Our device will contain the inducer for this inducible promoter which will cause the transcription of gene R and in-turn will inhibit the production of nucleases. So our constructs will be stable in our device. In case of plasmid getting lost into the environment, the gene R won’t be induced and thus will cause the expression of nucleases which will digest the DNA of the host organism including the detection device, thereby ensuring the biosafety of the device. As of now a possible candidate for R gene is Monococcal nuclease. It will be under the control of p(TetR) promoter. TetR coding sequence will be expressed under the control of pLac promoter which is induced by IPTG.

Expert Advice

One of our private sponsors is Lupin Limited which is a leading pharmaceutical company. We visited Lupin Research Park in Pune in order to collect expert advice in improving the safety of our module. We gave a presentation describing our project, to some of the industry experts, researchers, doctors working in the field of TB research. They gave us valuable feedback on the feasibility and relevance of our project. We got positive criticism from them which helped us in looking at our project with a new perspective. They suggested that we should consider attenuating bacteria before making their replication faster as an added biosafety measure. They also talked about extrapulmonary tuberculosis. We took a feedback after the presentation which included the following questions and the response is represented in the form graphs.

From L-R each row, the questions corresponding to the graphs below are: