Team:Cardiff Wales
Welcome
Hi, and welcome to the Cardiff_Wales iGEM2016 team. As the first team from our country to enter iGEM, we proudly represent Wales on the international stage.
Our Problem
Inefficient diagnosis of STIs can have negative socioeconomic impacts. Point of care diagnostics is gradually making its way to the market in a bid for greater efficiency, more sensitive results, and lower costs. In fact, the first HIV home testing kit was legalised in the UK just last year. For further benefits, we propose a novel point of care diagnostic tool using a split luciferase and dCas 9 system.
Our Solution
Cardiif_Wales has developed a novel bioluminescence detection system for point-of-care diagnostic testing, termed Cas-Find. In this system a Streptococcus pyogenes dCas9 isoform codon optimised for Escherichia coli is fused to C- the or N- terminal fragments of a thermostable pH-tolerant Photinus pyralis luciferase mutant. The coexpression of sgRNA constructs targets these chimeric proteins to adjacent sequences, resulting in the reconstitution of luciferase activity and bioluminescence in the presence of luciferin. This constitutes the positive signal for DNA detection. We aimed to undertake a proof-of-concept study of this system using sgRNA targeted to E. coli 16S rRNA, describe both the of this system in vitro, and the optimum distance between sgRNA targets.
A strength of our project involves our Integrated Human Practices in which we document our interactions with experts in the ethical use of diagnostic kits, the rules surrounding the use of GM products as well as our involvement in numerous outreach activites.
