title: Smart vesicle kit for in situ monitoring of intracellular telomerase activity using a telomerase-responsive probe.
abstract: A smart vesicle kit was designed for in situ imaging and detection of cytoplasmic telomerase activity. The vesicle kit contained a telomerase primer (TSP) and a Cy5-tagged molecular beacon (MB) functionalized gold nanoparticle probe, which were encapsulated in liposome for intracellular delivery. After the vesicle kit was transfected into cytoplasm, the released TSP could be extended in the presence of telomerase to produce a telomeric repeated sequence at the 3' end, which was just complementary with the loop of MB assembled on probe surface. Thus, the MB was opened upon hybridization to switch the fluorescent state from "off" to "on". The fluorescence signal depended on telomerase activity, leading to a novel strategy for in situ imaging and quantitative detection of the cytoplasmic telomerase activity. The cytoplasmic telomerase activity was estimated to be 3.2 × 10(-11), 2.4 × 10(-11), and 8.6 × 10(-13) IU in each HeLa, BEL tumor and QSG normal cell, respectively, demonstrating the capability of this approach to distinguish tumor from normal cells. The proposed method could be employed for dynamic monitoring of the cytoplasmic telomerase activity in response to a telomerase-based drug, suggesting the potential application in discovery and screening of telomerase-targeted anticancer drugs.
authors.affiliation: State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University , Nanjing 210093, P. R. China.
authors.forename: Ruocan
authors.initials: R
authors.lastname: Qian
authors.forename: Lin
authors.initials: L
authors.lastname: Ding
authors.forename: Liwen
authors.initials: L
authors.lastname: Yan
authors.forename: Manfei
authors.initials: M
authors.lastname: Lin
authors.forename: Huangxian
authors.initials: H
authors.lastname: Ju
mesh_terms: D002232:Carbocyanines / Q000737:chemistry; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D057075:Enzyme Assays / Q000295:instrumentation*; D006046:Gold / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D053768:Metal Nanoparticles / Q000737:chemistry; D008856:Microscopy, Fluorescence; D015335:Molecular Probes / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D019098:Telomerase / Q000032:analysis* / Q000378:metabolism; D014162:Transfection
chemical_list: D002232:Carbocyanines; D008081:Liposomes; D015335:Molecular Probes; C085321:cyanine dye 5; D006046:Gold; D019098:Telomerase