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Microchimica Acta

, 186:81 | Cite as

Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters

  • Shi Ting Lee
  • Ruman Rahman
  • Kasturi Muthoosamy
  • Nur Aliana Hidayah Mohamed
  • XiaoDi Su
  • Saad Tayyab
  • Siu Yee NewEmail author
Original Paper
  • 113 Downloads

Abstract

A fluorogenic probe has been developed for determination of telomerase activity using chimeric DNA-templated silver nanoclusters (AgNCs). The formation of AgNCs was investigated before (route A) and after (route B) telomerase elongation reaction. Both routes caused selective quenching of the yellow emission of the AgNCs (best measured at excitation/emission wavelength of 470/557 nm) in telomerase-positive samples. The quenching mechanism was studied using synthetically elongated DNA to mimic the telomerase-catalyzed elongation. The findings show that quenching is due to the formation of parallel G-quadruplexes with a –TTA– loop in the telomerase elongated products. The assay was validated using different cancer cell extracts, with intra- and interassay coefficients of variations of <9.8%. The limits of detection for MCF7, RPMI 2650 and HT29 cell lines are 15, 22 and 39 cells/μL. This represents a distinct improvement over the existing telomeric repeat amplification protocol (TRAP) assay in terms of time, sensitivity and cost.

Graphical Abstract

A method was developed using chimeric DNA-templated silver nanoclusters to detect telomerase activity directly in cell extracts. The sensitivity of this new method outperforms the traditional TRAP assay, and without the need for amplification.

Keywords

Cancer probe AgNCs Biomarker Biosensor TRAP G-quadruplex Telomers MCF7 HT29 RPMI 2650 

Notes

Acknowledgements

The authors would like to acknowledge Dr. Eunice Ngai and Rayan Sabra for supplying MCF7 and HT29, respectively. This work is supported by Fundamental Research Grant Scheme (FRGS) for financial support via FRGS/2/2014/SG06/UNIM/02/1.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2018_3194_MOESM1_ESM.docx (155 kb)
ESM 1 (DOCX 155 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.School of Pharmacy, Faculty of ScienceUniversity of Nottingham MalaysiaSemenyihMalaysia
  2. 2.Children’s Brain Tumour Research Centre (CBTRC), School of Medicine, Queen’s Medical CentreUniversity of NottinghamNottinghamUK
  3. 3.Nanotechnology and Advanced Materials (NATAM), Faculty of EngineeringUniversity of Nottingham MalaysiaSemenyihMalaysia
  4. 4.Centre of Clinical Science, Faculty of DentistryUniversity of Technology MARA Sungai Buloh Campus, Jalan HospitalSelangorMalaysia
  5. 5.Institute of Materials Research and EngineeringSingaporeSingapore
  6. 6.Department of ChemistryNational University of SingaporeSingaporeSingapore
  7. 7.School of Science and EngineeringUniversity of the Sunshine CoastMaroochydore DCAustralia
  8. 8.Institute of Biological Sciences, Faculty of ScienceUniversity of MalayaKuala LumpurMalaysia

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