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| Type |
Poster Presentation |
| Area |
생명화학 |
| Room No. |
포스터발표장 |
| Time |
4월 21일 (금요일) 13:00~14:30 |
| Code |
BIO.P-285 |
| Subject |
Detection of cell signaling molecules using live cell-based sensing systems |
| Authors |
전현진, 권영은*
동국대학교 의생명공학과, Korea
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| Abstract |
Biological signal molecules transmit information between cells in vivo and play key roles in modulating cell metabolism, functions, and development. Detecting signaling molecules in living cells is important for understanding their roles in biological systems and for studying various disease-related mechanisms. Herein, we describe a genetically encoded fluorescent sensor based on fluorescent trans-localization by intein-mediated reactions. The approach utilizes the generation of functional signal peptide by intein-mediated splicing or cleavage reaction which is activated in response to external stimuli. We first tested the concept of fluorescent trans-localization sensor by using a pair of split-inteins which react spontaneously. Then a model system was prepared to monitor the rapamycin induced hetero-dimerization of FKBP and FRB. Additionally, we constructed a live cell based sensing system to monitor the influx of Ca2+, an important intracellular signaling molecules related to various cellular events, using calmodulin and calmodulin binding domain conjugated split-intein system. The sensing system successfully reported the change of intracellular Ca2+ levels induced by ionophore or histamine. Finally, the sensing of corticosterone, which affects various physiological responses such as gluconeogenesis, deposition of hepatic glycogen, and anti-inflammatory response, was also carried out using glucocorticoid receptor protein. This self-replicable live cell-based sensing systems can be engineered to report various cellular events of interest in biologically relevant environment thus can be used for drug screening and disease mechanism study. |
| E-mail |
9212jhj@gmail.com |
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119th General Meeting of the KCS