Deaths of lung Cancer worldwide
What is the solution?
![](https://static.igem.wiki/teams/4170/wiki/home/prevent.png)
“Κάλλιον το προλαμβάνειν ή το θεραπεύειν”,
“Prevention is better than Cure”,
Our solution
Think out of the box to create the box!
![](https://static.igem.wiki/teams/4170/wiki/home/dias.png)
DIAS
Diagnostic Investigation Accurate System
for the early detection of lung cancer
Features
As scientists dedicated to cancer research, we focused our efforts in designing and developing an innovative in vitro diagnostic device for the detection of lung cancer at the early stages of the disease. Since lung cancer is one of the leading medical societal challenges today, we hope that our diagnostic device could be a real contribution to the medical society with grant social impact. Our diagnostic device is called DIAS and this name derives from Greek mythology and especially from the greek chief deity of the pantheon Zeus-Dias.
DIAS detection platform is a CRISPR/Cas13a-based molecular detection platform for the early detection of lung cancer. Our system's implementation combines knowledge from different scientific multidisciplinary fields such as biochemistry, molecular biology and microfluidics.
Our detection platform is based on the proper “assembly” and adeptly orchestrated function of 2 fundamental components:
The combination of these state-of-the-art techniques coming from different scientific fields can give the desire result: a rapid, cost-effective and highly sensitive diagnostic platform for lung cancer detection.
From the early stages of lung cancer development, some miRNAs exhibit elevated levels in the blood of patients compared to healthy people. These miRNA biomarkers are the “prey” of the CRISPR/Cas13a molecular system. The CRISPR/Cas13a system consists of the Cas13a “predator” and the crRNA specifically-designed molecules which are the “informers” and guide the Cas13 enzyme to the target miRNA. As soon as the “predator” finds the “prey”, the CRISPR/Cas13a system is activating, cutting off any RNA molecule nearby. Exploiting the cleavage activity of the Cas13a “molecular scissor”, we add to the reaction's environment an RNA reporter molecule or else “flashlight” which releases fluorescent signals after cleavage.
Thus, we provide our system's output with two possible options. The first one is the “ON” effect, where the patient's sample is enriched with the miRNA biomarker. Under these circumstances, the crRNA “informer” will hybridize with the miRNA “prey”, the Cas13a “predator” will be activated and the RNA reporter “flashlight” will be chopped emitting fluorescent signals.
The second possibility is the “OFF” effect in which the patient's sample contains relatively low amounts of the target miRNA. In this case the “informer” cannot perceive the miRNA “prey” and the Cas13 “predator” will remain inactive and no fluorescence will be emitted.
To enhance the system's sensitivity we decided to transfer this enzyme “predator-prey game” in small picoliter-sized droplets generated by droplet microfluidics. Minimizing the reaction volume or else the “game area” the Cas13a predator with the guide of the crRNA “informer” could easily detect the miRNA molecules even if these molecules are presented in low quantity in the sample. Therefore with the microfluidics chip implementation we can achieve ultrasensitive detection performance, simplifying the “translational process” of our diagnostic device towards implementation to the clinical practice.
How Does it Work?
In the real world...
In the gaming world...
Our Accomplishments
Synthetic biology-based methodologies for reproducible synthesis of any crRNA with a customized method (crRNA Preparation crPrep kit contribution page). | |
A bioinformatic analysis utilizing the R programming language to validate the candidate miRNA biomarkers in clinical samples derived from publicly available databases (bioinformatic analysis page). | |
The modeling of our project's biological components, the droplet microfluidics in silico modeling and the model-driven feedback to our experiments (model page). | |
The Meta-CRISPR part collection. A standardized and interoperable CRISPR/crRNA collection with wide applications in synthetic biology (best part collection). | |
The efficient production and purification of all the molecular components of our system such as the Cas13a and the crRNAs (results page). | |
A detailed CRISPR-based measurement workflow designed in a way to enable easy completion of the experimental procedures minimizing the influence of any involved factors that could disrupt system's reliability (measurement page). |
An extensive characterization of the Cas13/crRNA reaction conditions useful for every future iGEM team (engineering success, results). | |
Proof of our system's functionality to detect the target miRNA in complex biological matrices with high sensitivity (proof-of-concept page). | |
Droplet microfluidic chip manufacturing via 3D-printing technology (engineering page). | |
Standardization of the conditions, the flow rates and the reaction's volumes for the microfluidics fluorescence assay (proof-of-concept page). | |
Proof our system's functionality in picoliter-sized droplets generated by the microfluidic chips (proof-of-concept). | |
Well-characterized registry pages of all our genetic parts with useful information for the synthetic biology community (iGEM registry). |