121st General Meeting of the KCS

Type Poster Presentation
Area Life Chemistry
Room No. Event Hall
Time 4월 20일 (금요일) 11:00~12:30
Code BIO.P-311
Subject Genetically encoded biosensors to detect signaling molecules in living cells
Authors Hyunjin Jeon, Youngeun Kwon*
Department of Biomedical Engineering (BK21 plus), Dongguk University, Korea
Abstract Live cell-based biosensors have emerged as a useful tool for biotechnology and chemical biology. Genetically encoded sensor cells often utilize bimolecular fluorescence complementation or fluorescence resonance energy transfer to build a reporter unit that suffers from non-specific signal activation at high concentrations. Here, we designed genetically encoded sensor cells that can report the presence of biologically active molecules via fluorescence-translocation based on split intein-mediated conditional protein trans-splicing (PTS) and conditional protein trans-cleavage (PTC) reactions. In this work, the target molecules or the external stimuli activated intein-mediated reactions, which resulted in activation of the fluorophore-conjugated signal peptide. This approach fully valued the bond-breaking features of intein-mediated reactions in sensor construction and thus eliminated the interference of false-positive signals resulting from the mere binding of fragmented reporters. We could also avoid the necessity of designing split reporters to refold into active structures upon reconstitution. These live cell-based sensors were able to detect biologically active signaling molecules, cortisol, as well as relevant biological stimuli, such as the cortisol agonist Dexamethasone. These live cell-based sensing systems hold large potential for applications such as drug screening and toxicology studies, which require functional information about targets.
E-mail 9212jhj@gmail.com