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|Title:||GENETICALLY ENCODED FLUORESCENT PROTEIN AND CELL SENSORS FOR NANOTOXICITY STUDIES||Authors:||YOU FANG||Keywords:||Silver nanoparticles, Trojan-horse, intracellular dissolution of nanoparticles, silver sensor, FRET||Issue Date:||28-Jul-2016||Citation:||YOU FANG (2016-07-28). GENETICALLY ENCODED FLUORESCENT PROTEIN AND CELL SENSORS FOR NANOTOXICITY STUDIES. ScholarBank@NUS Repository.||Abstract:||Silver nanoparticles (AgNPs) are one of the most widely used nanomaterials with many applications, and concerns on AgNP-induced cytotoxic effects need to be carefully addressed prior to further use. However, existing data on AgNP-induced toxicity are debatable as it is hard to distinguish the toxic effects between AgNPs and ionic (Ag+) forms. In this thesis, genetically encoded fluorescent protein Ag+ sensors were developed for AgNP toxicity studies. First, a sensor namely MT2a-FRET was developed with good sensitivity and selectivity toward Ag+ but inert to AgNPs. By genetically targeting MT2a-FRET to two model organisms, the intracellular AgNP dissolutions were monitored. Results showed that AgNPs displayed “Trojan-horse” mechanism in mammalian cells, while the antibacterial effect of AgNPs was caused by the released Ag+ fraction from AgNPs in the medium. Finally, an improved sensor namely CαR-N2 was generated with better sensitivity and pH resistance, which was suitable for working in environmental relevant conditions.||URI:||http://scholarbank.nus.edu.sg/handle/10635/135259|
|Appears in Collections:||Ph.D Theses (Open)|
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