Antibiotic resistance is one of the serious issues in healthcare worldwide. There is a growing need of developing a simple and rapid method for sensitive detection of antibiotic resistant bacteria, which can lead to the prevention and earlier treatment of infectious diseases caused by antibiotic resistant bacteria. Transcriptional Activator-Like Effector (TALE) is a novel class of DNA-binding proteins with the unique modularity, flexibility and easy programmability. TALEs can be designed to bind to any DNA sequences through unique repeat variable di-residues (RVDs) present in each repeat in the central DNA-binding region of the protein. Graphene oxide (GO) is a 2D-nanosheet that has many oxygen-containing functional groups such as carboxyl, hydroxyl, carbonyl and epoxide in its structure. By taking advantage of the unique quenching ability of GO, we designed a novel detection system for screening and detecting antibiotic resistance gene (ARG) in bacteria utilizing engineered TALEs and GO as a novel sensing platform. In the absence of target DNA, fluorescence signal of quantum dot (QD)-labeled TALEs is quenched via fluorescence resonance energy transfer (FRET) between QDs and GO when they are in close proximity. In the presence of target DNA, the bound complex of TALEs and DNA is dissociated from GO, thereby restoring the fluorescence signal. In this study, we have demonstrated the novel application of a new class of DNA-binding protein TALEs and 2D nanosheet GO as a new sensing platform for detecting ARG in a simple and rapid assay.

Novel application of TALE and graphene oxide based biosensor for rapid detection of antibiotic resistance genes

ACS Fall 2021

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.