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; Xuai Du Su; Saad Tayyab; Siu Yee New

doi:10.1007/s00604-018-3194-7

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

Journal article

Peer reviewed

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, Xuai Du Su, Saad Tayyab and Siu Yee New

Microchimica Acta, Vol.186(81)

2019

DOI: https://doi.org/10.1007/s00604-018-3194-7

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Abstract

cancer probe

AgNCs

biomarker

biosensor

TRAP

G-quadruplex

telomers

MCF7

HT29

RPMI 2650

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 telomerasecatalyzed 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.

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Title: Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters
Authors: Shi Ting Lee (Author) - University of Nottingham, Malaysia
Ruman Rahman (Author) - University of Nottingham, Malaysia
Kasturi Muthoosamy (Author) - University of Nottingham, Malaysia
Nur Aliana Hidayah Mohamed (Author) - Jalan Hospital, Malaysia
Xuai Du Su (Author) - University of the Sunshine Coast
Saad Tayyab (Author) - University of Malaya, Malaysia
Siu Yee New (Author) - University of Nottingham, United Kingdom
Publication details: Microchimica Acta, Vol.186(81); 10
Publisher: Springer Wien
Date published: 2019
DOI: 10.1007/s00604-018-3194-7
ISSN: 0026-3672
Organisation Unit: University of the Sunshine Coast, Queensland
Language: English
Record Identifier: 99450764702621
Output Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Web Of Science research areas: Chemistry, Analytical

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