We had devoted ourselves to constructing devices to detect microRNA expression levels in a novel way that could be used for Alzheimer's disease (AD) diagnoses. Anti-microRNA-34c (BBa_K1940000) targeting miR-34c was a basic part which was applied in device microRNA-34c detector 1.0 and 2.0. Interestingly, our results showed device 1.0 (BBa_K1940001) could not only determine the existence of microRNA but also the quantity of microRNA. Further, we tested device 2.0 (BBa_K1940004- BBa_K1940011) which aim to make diagnoses easier and more directly and they also worked well.
Anti- microRNA-34c can be used as a miR-34c detector. In this part, we designed 4 series antisense strand of miR-34c which were linked by 4 linkers. Anti- microRNA-34c is sensitive to the expression levels of miR-34c. And combined with a reporter, we can diagnose AD accurately and rapidly (Fig. 1).
BBa_K1940001: microRNA-34c detector 1.0
We designed this part to detect the abundance of microRNA-34c. Anti-microRNA-34c was insert after a constitutive promoter which express GFP (Fig. 2). As anti- microRNA-34c is sensitive to the expression levels of miR-34c, we would have a direct impact on GFP’s expression. Determining GFP’s fluorescence intensity differences will justify the content amount of microRNA-34c. More experimental details, please visit results
BBa_K1940004---BBa_K1940011: microRNA-34c detector 2.0
In the system 2.0, we want to achieve the positive relationship of the fluorescent intensity and the concentration of miRNA. We had tried two NOT logic gate, tetR-Ptet and lacI-Plac, with the composition of different strength constitutive promoters (Fig. 3). More experimental details, please visit results