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| Type |
Poster Presentation |
| Area |
Life Chemistry |
| Room No. |
Event Hall |
| Time |
4월 20일 (금요일) 11:00~12:30 |
| Code |
BIO.P-307 |
| Subject |
Fluorescent biosensors for monitoring of signal molecules using signal peptide reconstitution |
| Authors |
Hyunjin Jeon, Jea Hee Ryu, Youngeun Kwon*
Department of Biomedical Engineering (BK21 plus), Dongguk University, Korea
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| Abstract |
Biological signal molecules are substances that transmit information in vivo and are involved in various actions such as modulate cellular metabolism, function, 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 that can be used as a new tool with general applicability for monitoring signaling molecules based on fluorescent trans-localization by intein mediated reactions. The approach utilizes the generation of functional signal peptide by splicing or cleavage 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 by histamine. This replicable live cell based sensing systems can be engineered to report various cellular events of interest. Also it provides biologically relevant information to be applied for drug screening and disease mechanism study.
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| E-mail |
9212jhj@gmail.com |
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121st General Meeting of the KCS