A three-dimensional DNA walker amplified FRET sensor for detection of telomerase activity based on the MnO<sub>2</sub>nanosheet-upconversion nanoparticle sensing platform

Xu, Mingdi; Zhuang, Junyang; Jiang, Xiaoyu; Liu, Xinzhong; Tang, Dianping

doi:10.1039/c9cc05387e

Cited by 55 publications

(27 citation statements)

References 33 publications

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“…Recently, different methods and strategies of detection of telomerase activity have been reported 48,49 . Compared with the traditional methods, the advantages of biotin‐labeled primer‐based nonradioactive detection method are simple operation, reliable results, accurate quantification, and easy promotion in ordinary laboratories.…”

Section: Discussionmentioning

confidence: 99%

Nonradioactive direct telomerase activity detection using biotin‐labeled primers

Wang

Jin

et al. 2021

Clinical Laboratory Analysis

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Background Telomerase is a ribonucleoprotein enzyme responsible for maintenance of telomere length which expressed in more than 85% of cancer cells but undetectable in most normal tissue cells. Therefore, telomerase serves as a diagnostic marker of cancers. Two commonly used telomerase activity detection methods, the telomerase repeated amplification protocol (TRAP) and the direct telomerase assay (DTA), have disadvantages that mainly arise from reliance on PCR amplification or the use of an isotope. A safe, low‐cost and reliable telomerase activity detection method is still lacking. Method We modified DTA method using biotin‐labeled primers (Biotin‐DTA) and optimized the method by adjusting cell culture temperature and KCl concentration. The sensitivity of the method was confirmed to detect endogenous telomerase activity. The reliability was verified by detection of telomerase activity of published telomerase regulators. The stability was confirmed by comparing the method with TRAP method. Results Cells cultured in 32°C and KCl concentration at 200 mM or 250 mM resulted in robust Biotin‐DTA signal. Endogenous telomerase activity can be detected, which suggested an similar sensitivity as DTA using radioactive isotope markers. Knockdown of telomerase assembly regulator PES1 and DKC1 efficiently reduced telomerase activity. Compared with TRAP method, Biotin‐DTA assay offers greater signal stability over a range of analyte protein amounts. Conclusion Biotin‐labeled, PCR‐free, and nonradioactive direct telomerase assay is a promising new method for the easy, low‐cost, and quantitative detection of telomerase activity.

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Section: Discussionmentioning

confidence: 99%

Nonradioactive direct telomerase activity detection using biotin‐labeled primers

Wang

Jin

et al. 2021

Clinical Laboratory Analysis

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“…A potential disadvantage in this context is that UCNPs are often larger than the ∼10 nm range of FRET. Energy transfer efficiencies are thus highest for emitters at the surface of the NP host and potentially negligible for emitters at the center. − One strategy for mitigating this challenge has been the use of Au NPs − and nanosheets , as more efficient energy acceptors. In one recent example, Au NPs and UCNPs were the corners of a DNA tetrahedron, where the Au NPs quenched the UCNP PL via energy transfer .…”

Section: Chemical Analysis and Bioanalysismentioning

confidence: 99%

Photoluminescent Nanoparticles for Chemical and Biological Analysis and Imaging

et al. 2021

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Research related to the development and application of luminescent nanoparticles (LNPs) for chemical and biological analysis and imaging is flourishing. Novel materials and new applications continue to be reported after two decades of research. This review provides a comprehensive and heuristic overview of this field. It is targeted to both newcomers and experts who are interested in a critical assessment of LNP materials, their properties, strengths and weaknesses, and prospective applications. Numerous LNP materials are cataloged by fundamental descriptions of their chemical identities and physical morphology, quantitative photoluminescence (PL) properties, PL mechanisms, and surface chemistry. These materials include various semiconductor quantum dots, carbon nanotubes, graphene derivatives, carbon dots, nanodiamonds, luminescent metal nanoclusters, lanthanidedoped upconversion nanoparticles and downshifting nanoparticles, triplet−triplet annihilation nanoparticles, persistent-luminescence nanoparticles, conjugated polymer nanoparticles and semiconducting polymer dots, multi-nanoparticle assemblies, and doped and labeled nanoparticles, including but not limited to those based on polymers and silica. As an exercise in the critical assessment of LNP properties, these materials are ranked by several application-related functional criteria. Additional sections highlight recent examples of advances in chemical and biological analysis, point-of-care diagnostics, and cellular, tissue, and in vivo imaging and theranostics. These examples are drawn from the recent literature and organized by both LNP material and the particular properties that are leveraged to an advantage. Finally, a perspective on what comes next for the field is offered.

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“…Its advantages are significant Stokes shift, narrow emission spectrum, long lifetime, etc. Compared with downconversion conventional fluorescent probes, UCNPs reduce the background of autofluorescence and enhances light stability, significantly improving the signal-to-noise ratio in biological detection based on fluorescent labels. − Consequently, UCNPs hold great promise for biodetection based on fluorescence labeling. It is well known that the efficiency of FRET is highly dependent on the distance between the donor and the acceptor, and nonradiative energy transfer is usually effective within a distance of 10 nm .…”

Section: Introductionmentioning

confidence: 99%

Upconversion Fluorescence Resonance Energy Transfer Aptasensors for H5N1 Influenza Virus Detection

Zhao

Wang

et al. 2021

ACS Omega

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Influenza A virus (IAV) poses a significant threat to human health, which calls for the development of efficient detection methods. The present study constructed a fluorescence resonance energy transfer (FRET) system based on novel fluorescent probes and graphene oxide (GO) for detecting H5N1 IAV hemagglutinin (HA). Here, we synthesized small (sub-20 nm) sandwich-structured upconversion nanoparticles (UCNPs) (SWUCNPs for short) with a high energy transfer efficiency, which allows for controlling the emitter in a thin shell. The π–π stacking interaction between the aptamer and GO shortens the distance between the fluorescent probe and the receptor, thereby realizing fluorescence resonance energy transfer (FRET). When HA is present, the aptamer enables changes in their conformations and move away from GO surface. Fluorescence signals display a linear relationship between HA quantitation in the range of 0.1–15 ng mL –1 and a limit of detection (LOD) of 60.9 pg mL –1 . The aptasensor was also applicable in human serum samples with a linear range from 0.2 to 12 ng mL –1 and a limit of detection of 114.7 pg mL –1 . This strategy suggested the promising prospect of the aptasensor in clinical applications because of the excellent sensing performance and sensitivity. This strategy may be promising for vitro diagnostics and provides new insights into the functioning of the SWUCNPs system.

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A three-dimensional DNA walker amplified FRET sensor for detection of telomerase activity based on the MnO₂nanosheet-upconversion nanoparticle sensing platform

Abstract: We report a fluorescent sensing platform for telomerase activity assay by coupling a 3D DNA walker with the MnO2-UCNPs-based FRET system.

Cited by 55 publications

References 33 publications

Nonradioactive direct telomerase activity detection using biotin‐labeled primers

Nonradioactive direct telomerase activity detection using biotin‐labeled primers

Photoluminescent Nanoparticles for Chemical and Biological Analysis and Imaging

Upconversion Fluorescence Resonance Energy Transfer Aptasensors for H5N1 Influenza Virus Detection

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A three-dimensional DNA walker amplified FRET sensor for detection of telomerase activity based on the MnO2nanosheet-upconversion nanoparticle sensing platform

Abstract: We report a fluorescent sensing platform for telomerase activity assay by coupling a 3D DNA walker with the MnO2-UCNPs-based FRET system.

Cited by 55 publications

References 33 publications

Nonradioactive direct telomerase activity detection using biotin‐labeled primers

Nonradioactive direct telomerase activity detection using biotin‐labeled primers

Photoluminescent Nanoparticles for Chemical and Biological Analysis and Imaging

Upconversion Fluorescence Resonance Energy Transfer Aptasensors for H5N1 Influenza Virus Detection

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Resources

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A three-dimensional DNA walker amplified FRET sensor for detection of telomerase activity based on the MnO₂nanosheet-upconversion nanoparticle sensing platform