ENZYME-SWITCHES · Enzyme-conjugated DNA nanoswitches for biosensing applications

Enzyme-conjugated DNA nanoswitches for biosensing applications

The present fellowship proposed the development of a new generation of enzyme-based biosensors for diagnostic applications. I proposed to use enzymes conjugated with synthetic DNA responsive nanoswitches, with the main goal of combining the high specificity of enzymes and the high programmability and versatility of DNA nanodevices, According to the general idea, the enzyme will recognize a specific diagnostic target and will catalyze the formation of a product. Then the DNA responsive nanoswitch will be able to recognize and detect the enzymatic product. As we have previously reported, the direct conjugation among a single enzyme and a single DNA nanoswitch could create issues in obtaining good results. Thus, in order to improve the communication among enzymes and DNA modules, we decided to study the possibility of using a polymeric DNA-based structure as a scaffold to co-localize biological macromolecules (i.e. enzymes) with several DNA-responsive modules (i.e. DNA nanoswitches). More specifically, to do that we have employed the well-known DNA tubular structures, tile-based DNA structures obtained through the self-assembly of double-cross-over DNA tiles (DAE-E). As already demonstrated, it is also possible to re-engineer different tiles with multiple functional groups and the same sticky ends, so that they can co-assemble and form a unique DNA scaffold with multiple functionalities Nowadays, biosensors have become important devices in diagnosing and monitoring health. Among them, enzyme-based biosensors are one of the most successful examples, but they are still limited to measure few clinical targets. So the development of new strategies to create versatile platforms able to detect a wide range of clinical targets is needed. To achieve this challenging purpose, several objectives have been defined: • Design and optimization of the several pH-dependent DNA-based nanodevices. • Study and characterization of aptamer-based DNA nanodevices. • Synthesis and optimization of the enzyme-conjugated DNA nanoswitches. • Design and optimization of the DNA scaffold to co-localize the enzyme and the reported DNA-nanodevice. • Development of novel strategies to dynamically introduce functional groups in DNA structures. • Characterization of the strand displacement reaction, one of the most used reaction to control the assembly of DNA structures. • Demonstration of the orthogonality and versatility of the proposed strategies. • Demonstration of the versatility of the proposed approach employing other enzyme-based biosensors.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз