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Genetically encoded biosensors for the detection of rapamycin: toward the screening of agonists and antagonists.
Analyst. 2020 Jul 03 [Online ahead of print]A

Abstract

Biosensors are valuable tools for the rapid screening of biological targets with high sensitivity and specificity. It is important to screen biological events in their native context for pharmacological and toxicological applications. However, in vitro biosensors often require purified probes and targets for screening, thus providing limited information on the biological activities of targets in their native environment. To address this issue, we developed a cell-based sensing system that could detect a biologically active small molecule, rapamycin (Rapa). We designed a reporter system based on fluorescence translocation by signal peptide reconstitution. Herein, signal peptides are activated by conditional protein splicing without the need for refolding into a functional tertiary structure, thus eliminating false positives and negatives due to mere binding or misfolding. The developed biosensor demonstrated excellent sensitivity with a limit of detection of 0.1 nM, and it was able to screen the agonist and antagonist of Rapa. The developed cell-based sensing system could contribute to improving the screening system aimed to identify the natural mimetics of Rapa and potential drug candidates.

Authors+Show Affiliations

Department of Biomedical Engineering (BK21 plus), Dongguk University, Seoul 04620, Korea. ykwon@dongguk.edu.Department of Biomedical Engineering (BK21 plus), Dongguk University, Seoul 04620, Korea. ykwon@dongguk.edu and Gencurix Inc. 3F, 242 Digital-ro, Guro-gu, Seoul 08394, Korea.Department of Biomedical Engineering (BK21 plus), Dongguk University, Seoul 04620, Korea. ykwon@dongguk.edu.Department of Biomedical Engineering (BK21 plus), Dongguk University, Seoul 04620, Korea. ykwon@dongguk.edu.Department of Biomedical Engineering (BK21 plus), Dongguk University, Seoul 04620, Korea. ykwon@dongguk.edu.Department of Biomedical Engineering (BK21 plus), Dongguk University, Seoul 04620, Korea. ykwon@dongguk.edu.Department of Biomedical Engineering (BK21 plus), Dongguk University, Seoul 04620, Korea. ykwon@dongguk.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32618311

Citation

Lee, Euiyeon, et al. "Genetically Encoded Biosensors for the Detection of Rapamycin: Toward the Screening of Agonists and Antagonists." The Analyst, 2020.
Lee E, Jeon H, Ryu J, et al. Genetically encoded biosensors for the detection of rapamycin: toward the screening of agonists and antagonists. Analyst. 2020.
Lee, E., Jeon, H., Ryu, J., Kang, C., Kim, S., Park, S., & Kwon, Y. (2020). Genetically encoded biosensors for the detection of rapamycin: toward the screening of agonists and antagonists. The Analyst. https://doi.org/10.1039/d0an01116a
Lee E, et al. Genetically Encoded Biosensors for the Detection of Rapamycin: Toward the Screening of Agonists and Antagonists. Analyst. 2020 Jul 3; PubMed PMID: 32618311.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Genetically encoded biosensors for the detection of rapamycin: toward the screening of agonists and antagonists. AU - Lee,Euiyeon, AU - Jeon,Hyunjin, AU - Ryu,Jeahee, AU - Kang,Chungwon, AU - Kim,Soyoun, AU - Park,Seungil, AU - Kwon,Youngeun, Y1 - 2020/07/03/ PY - 2020/7/4/entrez JF - The Analyst JO - Analyst N2 - Biosensors are valuable tools for the rapid screening of biological targets with high sensitivity and specificity. It is important to screen biological events in their native context for pharmacological and toxicological applications. However, in vitro biosensors often require purified probes and targets for screening, thus providing limited information on the biological activities of targets in their native environment. To address this issue, we developed a cell-based sensing system that could detect a biologically active small molecule, rapamycin (Rapa). We designed a reporter system based on fluorescence translocation by signal peptide reconstitution. Herein, signal peptides are activated by conditional protein splicing without the need for refolding into a functional tertiary structure, thus eliminating false positives and negatives due to mere binding or misfolding. The developed biosensor demonstrated excellent sensitivity with a limit of detection of 0.1 nM, and it was able to screen the agonist and antagonist of Rapa. The developed cell-based sensing system could contribute to improving the screening system aimed to identify the natural mimetics of Rapa and potential drug candidates. SN - 1364-5528 UR - https://www.unboundmedicine.com/medline/citation/32618311/Genetically_encoded_biosensors_for_the_detection_of_rapamycin:_toward_the_screening_of_agonists_and_antagonists L2 - https://doi.org/10.1039/d0an01116a DB - PRIME DP - Unbound Medicine ER -
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