Team:CMUQ/Description
Project Description
The project at a glance:
How it all started..
In a world where healthcare and health insurance is a primary concern for citizens and policymakers, we saw a need to create an equal opportunity for one of the most important and relatively expensive tests worldwide.
Testing for genetic disorders, although taken for granted in some countries, is a major gap in others. Whether it is the lack of awareness, equipment, expertise or simply the overpriced exams, the Middle East and many other less fortunate regions suffer from the uncontrolled spread of genetic disease. We found no better way to describe our motives than by collectively extrapolating our drives to take part in this research. We believe that combining our different cultural backgrounds, skills, knowledge, and passions enabled us to try and improve the lives of those in need, whilst also contributing to the scientific community. Lastly, our trust in the essentiality of the technique we are introducing allowed us to participate again with a developed vision of our last iGEM idea.
What inspired us?
Inspired by Dr. Doudna’s published work “CRISPR- Cas12a target binding unleashes indiscriminate single-stranded DNase activity”, we are utilizing CRISPR for purposes other than gene editing to create a novel, field-ready diagnostic technique for carriers of recessive traits. Cas12a proteins are DNA targeting enzymes that recognize DNA based on a guide RNA (gRNA) sequence designed to match a target. The binding initiates non-specific single-stranded DNA (ssDNA) cleavage activity in Cas12a, sufficient to degrade linear and circular ssDNA within minutes. Through this, ssDNA attached to fluorescent dye and quencher, serving as reporters, will undergo degradation. Upon cleavage, the quencher is released and fluorescence is emitted.
Elaborating further, our mechanism includes a gRNA that is complementary to a specific SNP causing a recessive genetic disease, which will be integrated into the Cas12a protein such that when the target DNA is processed by Cas12a and bound to the gRNA, cleavage activity is initiated and fluorescence is emitted. Afterward, we use the device, which we built and programmed in-house, to detect this fluorescence with high sensitivity.
What is new this year?
Although the foundations of our research project were presented by our team in the iGEM 2018 round, this year we are integrating the judges’ and other expert feedback to optimize the sensitivity of our device, provide a pre-counseling prediction calculation and broaden our technique, allowing us to identify other similar genetic diseases. We plan to achieve this by measuring the fluorescence emitted from the Cas12a processing of a template DNA, comparing this to double-blinded DNA samples to verify our results. Moreover, we are designing gRNAs for locally and globally common genetic diseases that include Sickle Cell Anemia and Cystic Fibrosis. As for the technical portion of the project, the hardware and software teams are working on integrating machine learning, database analysis, and statistical evaluation of results into a software program that is bluetooth compatible with our improved hand-held device.
Biology + Technology coming together!
The combination of the biological technique, programmed software, and in-house designed device will yield both an on-field and in-lab genetic testing method that overcomes the cost and labor intensity limitations of sequencing. It will also eliminate the biohazardous use of blood by efficiently extracting DNA from saliva samples, making our technique less invasive. Altogether, our project introduces an eye-opening perspective on the broader uses of CRISPR in the field of synthetic biology.