"
Team:Fatih-Medical/Sherlocoli/Goals
From 2012.igem.org
It is aimed to put the earlier diagnosis by the detection of CTC and oncogenes present in blood before the metastasis phase of cancer. Two distinct pathways were contemplated for the detection of CTCs and oncogenes.
CTC – EpCAM
Our purpose is to catch the CTCs (Circulating Tumor Cells) which begin to circulate in blood in pre-metastasis stage of cancer via their EpCAM (Epithelial Cell Adhesion molecule) antigen sites. C215, one of the EpCAM specific Anti-EpCAMs, will be used for this. Antibodies, which will be fixed on the extracellular membrane of bacteria, are planned to bind to antigens of CTC. With the result of antigen-antibody merger, terminals of intracellular placed TEV protease will coalescent and activate the signal pathway; the head constituent of diagnosis implementation.
Free DNA
We will head for diagnosis by means of oncogene determination straying in the blood. This pathway is technically very similar to the CTC pathway; yet TALe (Transcription Activator Like effector) will be placed to extracellular matrix instead of the C215 antibodies. By the help of TALe; synthesized in conformity with sequence of oncogene’s determined region, oncogene will be caught and TEVp system underneath will shift to active unified condition from split state. TEVp-dependent manifold signaling pathways will be established for diagnosis.
For cancer detection the next major part working out after the CTC/oncogene capture is signaling.
Signaling
A signaling system, which will give required feedbacks for diagnosis after the cohesion of CTC/oncogene and designed system, was determined. In case of encounter with assorted situations we designed two workouts:
FRET
In this procedure, which stands on electrochemical system, Cerulean and Venus proteins will be utilized. These two color proteins will attach to each other by dint of TEV rec. site-owning chain. By application of light before the CTC/oncogene clutch we hope to obtain color from Venus whilst through the activation of TEVp after the capture these two will split up and we’ll try to acquire color from Cerulean.
Fast response
We decided to try out 2010 ICL’s fast response system as we expect the bacteria to work properly in circumferences like human blood. The goal is to optimize this system in surroundings like human blood as chemicals like catechol will be used and make comments through fast response signals.
In this procedure, which stands on electrochemical system, Cerulean and Venus proteins will be utilized. These two color proteins will attach to each other by dint of TEV rec. site-owning chain. By application of light before the CTC/oncogene clutch we hope to obtain color from Venus whilst through the activation of TEVp after the capture these two will split up and we’ll try to acquire color from Cerulean.
Fast response
We decided to try out 2010 ICL’s fast response system as we expect the bacteria to work properly in circumferences like human blood. The goal is to optimize this system in surroundings like human blood as chemicals like catechol will be used and make comments through fast response signals.
