Team:TU-Eindhoven/Part Collection
Parts
“The whole is more than the sum of its parts” - Aristotle
All parts used for IBDetection to be successful can be found on these pages. Part Overview gives a brief overview of all parts used in the sensor-reporter system and the Part Collection explains our used and designed collection in more detail, which is separated in the Basic Parts page and the Composite Parts page. Furthermore, the button Improvement brings you to the page of our improved part.
Part Collection
Worldwide, the incidence of IBD has risen steadily, and the exact cause of IBD is still unknown. Current methods for screening IBD patients are invasive, complex, unpleasant, expensive, and time-consuming (for more details check out Human Practices segment The Need). To decrease these negative features we designed a non-invasive monitoring method, which consists of GMOs containing a tetrathionate sensing-reporter cascade. Our designed system senses tetrathionate and in response produces gas vesicles, which can be detected using ultrasound equipment. Our part collection ranges from part BBa_K3972000 to part BBa_K3972006, which includes the various parts required for our system. We combined the sensing two-component system TtrS/R (BBa_K3972000 & BBa_K3972001) with ARG1 (BBa_K3972002) gas vesicle expression as output and demonstrated that these two can be joined together, which can be read about on our Proof of Concept page and Results page.
Composite part BBa_K3972003 and composite part BBa_K3972004 are both integrated in design A and design B respectively. The design A part consists of a TtrB185-269 promoter (BBa_K2507019), combined with the basic part ARG1. The design B part contains the same parts, with additional parts TtrR, TetR (BBa_K1745003) and pLtetO-1 promoter (BBa_K3332034) (Figure 1). The basic part TtrS functions as a transmembrane protein that binds tetrathionate resulting in activation and subsequent phosphorylation of TtrR. The kinase TtrR can bind the DNA as a phosphorylated dimer to induce gene expression. In both design A and design B, the TtrS/R two-component system is combined with the basic part ARG1. The ARG1 gene sequence codes for a collection of 12 proteins which assemble into gas vesicles upon expression. The expression of the ARG1 proteins in design A and design B are both controlled by pTtrB185-269. With our collection, we also prove that ARG1 as reporter system can be combined with different sensing systems, e.g. thiosulfate sensor, meaning it is possible to develop multiple types of sensors with gas vesicles as output.
For part improvement, composite part BBa_K3972005 was designed, which shows that the expression of proteins can be increased by using a 5’UTR region, additionally to the ribosome binding site (RBS). This indicates that our sensor, based on protein expression, could give higher expression levels.
All in all, our part collection demonstrates a (faster) tetrathionate sensor, followed by the expression of gas vesicles, which can be detected using ultrasound equipment. This system can give rise to a non-invasive monitoring method for IBD.
Figure 1: Overview part collection.