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Revision as of 12:46, 11 August 2016
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Project Description
There is no global challenge worth greater investment than improving quality of life. During initial discussions, we identified a wide range of problems that we felt further scientific development would be most beneficial: increasing crop yields; sewage filtration; reclamation of rare earth metals. Upon evaluation, the team decided that synthesizing a modular biosensor detection kit would be most influential, due to the potential application to a wide range of diseases and environmental health issues. This affordable, accessible system could tackle multiple issues over a large demographic, making the venture truly worthwhile. Lyme’s disease, although not at the forefront of discussion, is the inceptive focus of the investigation, as current detection methods can only be applied after debilitating symptoms have manifested. Our novel biosensor would therefore allow treatment of the disease at a period before serious symptoms have arisen. The theoretically adaptable nature of the bio detector also could allow replacement of antibody-reliant detection systems, hence why our other primary research focus is leptospirosis. We also aim to modify this technology such that it can be used to monitor toxic lead and mercury levels in water supplies.
Our detection system for infectious agents and environmental pollutants will be based on CRISPR/Cas9 technology, relying on conformational changes in an RNA-based sensor that will trigger transcriptional regulation by a dCas9 protein. We plan to produce a modular gene circuit that can detect the presence of either RNA from an infectious agent or metallic ion and output a fluorescent signal. This will be arranged on a plasmid encoding for an RNA sensor, the CRISPR/Cas9 system, and a fluorescent reporter. The RNA sensor will be a modified sgRNA containing a motif binding either Borrelia/Leptospira RNA or lead/mercury aptamers. This binding will cause a conformational change such that the modified dCas9 enzyme may bind upstream of the transcriptional start site of the fluorescent reporter gene inducing transcription. We will be testing this using multiple novel dCas9 fusion proteins as well as more traditional CRISPR/dCas9 methods. Afterwards, the sensor will be freeze-dried onto a paper scaffold for ease of use.
An example of a modified CRISPR/dCas9 system to be used in the project.
By the end of the project we hope to create a paper based sensor that will be able to be used in a low-tech, out-of-lab, environment. We hope this will make it available to less economically developed countries as a frontline diagnostic tool and make a real impact in the fight against infectious diseases and pollution.
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