Team:Brasil-SP

     Our project consists of a biological molecular device for detection of early stage chronic kidney disease using Cystatin C as biomarker and Bacillus subtilis as chassis. The genetic circuit being assembled is based on the outstanding project of iGEM 2010 Imperial College of London team (special thanks to the ex-iGEMer Christopher Hirst, who helped us a lot sending some important BioBricks). Part of our mission is also to improve characterization of the BioBricks developed in 2010 and to validate the molecular design as Cystatin C detection system. This flexibility of detection is based on a protease cleavage of a membrane protein which triggers the genetic circuit. Since any cleavage site could be designed, virtually any protease could be used as a signal for the detection. In our case, the disease biomarker will inhibit the action of Cathepsin S protease and the detection will be made indirectly and negatively - i.e. by the Cathepsin lack of protease activity and absense of the system output. We are on the way to assemble all the parts and properly characterize each part of our construction on time for the Jamboree.
     To address a real world situation, we are working on the same principle and aesthetics of the well known devices for biodetection like pregnancy or HIV tests: easy-to-use microfluidic devices. The plan is to design a microchip able to store spores of the developed strains of B. subtilis and safely expose blood samples to our biodetection system, successfully containing the biomaterial and enabling a proper discard of the chip. A priori, the device output monitoring would require a fluorescence detector tool, but we also propose a naked eye output observation as a concept for future prospects.
      Since we are working on a solution for a problem directly related to ordinary people, having a public feedback about synthetic biology is very important to analyze the social impact of our work and it help us to evaluate the biosafety and bioethical issues beyond a simple risk analysis - a sociological characterization of the values of our project. Thus, as a policy and practices approach, we will try to report Brazil's public opinion on these issues using a questionnaire to evaluate our actual scenario and, in a certain way, our own project.

In collaboration with the team Paris_Saclay, several abstracts were translated to different languages. The main objective is to divulgate the iGEM projects to the greatest number of people as possible.

Chronic Kidney Disease (CKD), which is characterized by alterations in kidney functions and structure, affects millions of people worldwide and a large portion of them is unaware of it. Absence of symptoms in early stages leads to a late diagnosis when patients need dialysis or even transplants. Currently, CKD is diagnosed by measuring creatinine levels in blood, which in turn are detectable only at late stages of renal dysfunction as well as are sensitive to factors such as diet, gender, ethnicity, age, muscle mass. We report the development of a biosensor that can diagnosis CKD in its early stages, identifying a biomarker named Cystatin C. Using a cell surface biosensor as detector and quorum-sensing system as transducer and response mechanism, we developed a genetic circuit that establishes a threshold, differentiating concentration ranges of Cystatin C. We envisioned it as a fast, simple and reliable tool for CKD screening and diagnosis.

A Doença Renal Crônica (DRC), caracterizada por alterações nas funções e na estrutura dos rins, afeta milhões de pessoas no mundo todo e grande porção delas não tem consciência disso. A ausência de sintomas em estágios iniciais leva a um diagnóstico tardio, quando os pacientes já necessitam de diálise ou até mesmo de transplantes. Atualmente, a DRC é diagnosticada através da quantificação de creatinina no sangue, que por sua vez é alterada a proporções detectáveis somente nos estágios mais avançados da disfunção renal, como também é sensível a fatores como: dieta, gênero, etnia, idade e massa muscular. Reportamos aqui o desenvolvimento de um biossensor que pode diagnosticar DRC em seus estágios iniciais, identificando um biomarcador chamado Cistatina C. Usando a membrana celular como detector e um sistema de quorum sensing como mecanismo de transdução e de resposta, desenvolvemos um circuito gênico que estabelece uma barreira e diferencia faixas de concentração de Cistatina C. Nós o idealizamos como uma ferramenta rápida, simples e confiável para o diagnóstico de DRC. (Translated by Brasil-SP Team)

L’insuffisance rénale chronique (CDK) est caractérisée par une altération de la fonction et de la structure du rein. Elle atteint des millions de personnes à travers le monde mais un grand nombre d’entre elles ignorent qu’elles en sont atteintes. L’absence de symptômes au cours des stades précoces retarde le diagnostique et les patients ont d’ores et déjà besoin de dialyse ou même d’une transplantation. Actuellement, le diagnostique repose sur le dosage sanguin de la créatinine qui n'est détectable qu'au stade de dysfonction rénale et est influencé par différents facteurs tels que le jeûne, le genre, l’origine ethnique, l’âge et la masse musculaire. Nous avons développé un biosenseur capable de détecter cette maladie à des stades précoces grâce à l’identification d’un biomarqueur appelé Cystatin C. En ajoutant un biosenseur à la surface des cellules comme détecteur et un système de quorum-sensing comme mécanisme de transduction et de réponse, nous avons développé un circuit génétique à seuil permettant la détection de différentes gammes de concentrations en Cystatin C. Nous pensons que cet outil permet un dépistage et un diagnostique de l’insuffisance rénale rapide, simple et fiable. (Translated by Paris_Saclay team)

Chronisch nierfalen, gekenmerkt door veranderingen in functie en structuur van de nier, treft miljoenen mensen wereldwijd en de meesten van hen zijn zich er niet van bewust. Een gebrek aan symptomen in vroege stadia van de ziekte leidt tot late diagnose wanneer patienten al dialyse of zelfs transplantatie nodig hebben. Momenteel wordt chronisch nierfalen gediagnostiseerd door het creatine niveau in het bloed te meten wat alleen meetbaar is in late stadia van nierfalen en gevoelig is voor factoren als dieet, geslacht, herkomst, leeftijd en spiermassa. We bespreken de ontwikkeling van een biosensor die nierfalen in vroege stadia kan diagnostiseren door de biomarker Cytostatin C te identiceren. Met een biosensor aan het oppervlak van de cel als detector en een quorum-sensing system als transductie en reactive mechanism hebben wij een genetisch circuit ontwikkeld dat een drempelwaarde vast kan stellen om te discrimineren tussen concentraties van Cytostatine C. Wij voorzien dat een snel, simpel en betrouwbaar gereedschap wordt voor screening en diagnose van nierfalen. (Translated by Groningen team)