Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters
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...
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| Format: | Article |
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Springer Verlag (Germany)
2019
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| _version_ | 1796960796930998272 |
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| author | Lee, Shi Ting Rahman, Ruman Muthoosamy, Kasturi Mohamed, Nur Aliana Hidayah Su, XiaoDi Tayyab, Saad New, Siu Yee |
| author_facet | Lee, Shi Ting Rahman, Ruman Muthoosamy, Kasturi Mohamed, Nur Aliana Hidayah Su, XiaoDi Tayyab, Saad New, Siu Yee |
| author_sort | Lee, Shi Ting |
| collection | UM |
| description | 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. |
| first_indexed | 2024-03-06T05:50:01Z |
| format | Article |
| id | um.eprints-20040 |
| institution | Universiti Malaya |
| last_indexed | 2024-03-06T05:50:01Z |
| publishDate | 2019 |
| publisher | Springer Verlag (Germany) |
| record_format | dspace |
| spelling | um.eprints-200402019-01-17T03:50:44Z http://eprints.um.edu.my/20040/ Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters Lee, Shi Ting Rahman, Ruman Muthoosamy, Kasturi Mohamed, Nur Aliana Hidayah Su, XiaoDi Tayyab, Saad New, Siu Yee Q Science (General) QH Natural history TA Engineering (General). Civil engineering (General) 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. Springer Verlag (Germany) 2019 Article PeerReviewed Lee, Shi Ting and Rahman, Ruman and Muthoosamy, Kasturi and Mohamed, Nur Aliana Hidayah and Su, XiaoDi and Tayyab, Saad and New, Siu Yee (2019) Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters. Microchimica Acta, 186 (2). p. 81. ISSN 0026-3672, DOI https://doi.org/10.1007/s00604-018-3194-7 <https://doi.org/10.1007/s00604-018-3194-7>. https://doi.org/10.1007/s00604-018-3194-7 doi:10.1007/s00604-018-3194-7 |
| spellingShingle | Q Science (General) QH Natural history TA Engineering (General). Civil engineering (General) Lee, Shi Ting Rahman, Ruman Muthoosamy, Kasturi Mohamed, Nur Aliana Hidayah Su, XiaoDi Tayyab, Saad New, Siu Yee Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters |
| title | Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters |
| title_full | Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters |
| title_fullStr | Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters |
| title_full_unstemmed | Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters |
| title_short | Amplification-free and direct fluorometric determination of telomerase activity in cell lysates using chimeric DNA-templated silver nanoclusters |
| title_sort | amplification free and direct fluorometric determination of telomerase activity in cell lysates using chimeric dna templated silver nanoclusters |
| topic | Q Science (General) QH Natural history TA Engineering (General). Civil engineering (General) |
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