DNAzyme-based probes for telomerase detection in early-stage cancer diagnosis.
Hui Wang,Michael J. Donovan,Ling Meng,Zilong Zhao,Zilong Zhao,Youngmi Kim,Mao Ye,Weihong Tan,Weihong Tan +8 more
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TLDR
A DNAzyme-based probe to detect the presence of telomerase in cell lysates with a fivefold increase in signal level for cancer cells over normal cells is described.Abstract:
Human telomerase is a polymerase enzyme that adds tandem repeats of DNA (TTAGGG) in the telomeric region to the ends of chromosomes. Since telomerase can be detected in immortalized, but not normal, somatic cells, it has been considered a selective target for cancer chemotherapy. Here, we describe a DNAzyme-based probe to detect the presence of telomerase in cell lysates. Telomerase elongates the primer site on the probe. Subsequent addition of the Pb(II) cofactor activates the DNAzyme, which cleaves the elongated fragment at the RNA site, releasing the probe for repetitive cycling and signal amplification. The cleaved fragment is detected by a reporter molecular beacon. Enzymatic amplification with rapid turnover allows detection of telomerase in the range of 0.1 to 1 μg cell lysate, with a fivefold increase in signal level for cancer cells over normal cells. This probe design can provide a simple, yet rapid and sensitive, measurement of telomerase activity.read more
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References
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Journal Article
Telomerase activity in premalignant and malignant lesions of human oral mucosa.
S. Kannan,Hidetoshi Tahara,Hiroshi Yokozaki,Babu Mathew,K R Nalinakumari,M.K. Nair,Eiichi Tahara +6 more
TL;DR: It is shown that telomerase activity is present in normal oral squamous epithelium, as it is in normal hematopoietic cells and in carcinomas, and that telomersase activity has a relationship with degree of tumor differentiation and treatment response.
Journal ArticleDOI
Telomerase Activity in Human Urothelial Tumors
TL;DR: Findings indicate that telomerase activation may be a critical step in the pathogenesis of urothelial tumors and no significant correlation was observed between high levels of telomersase expression and the clinicopathologic features of the tumors, including clinical stage, pathologic grade, tumor multiplicity, and status of recurrence.
Journal ArticleDOI
Detection of telomerase activity by combination of trap method and scintillation proximity assay (spa).
TL;DR: The adaptation of conventional TRAP assay to SPA is described and the possibility of using this system for high-throughput screening for screening large number of compounds or clinical samples is shown.
Journal ArticleDOI
Detection of activity of telomerase in tumor cells using fiber optical biosensors
TL;DR: A new biosensoric assay based on total internal reflection fluorescence measuring the activity of the telomerase on sensor surface shows with the modified biosensor that the PS/TS primer binds the telomersase from the HL-60 cell lysates, effectively elongating the immobilized primer.
Journal ArticleDOI
New method to measure telomerase activity by transcription-mediated amplification and hybridization protection assay
TL;DR: It is demonstrated that the modified TMA can detect and measure telomerase activity and that TMA/HPA is as sensitive and reproducible as conventional TRAP, but is both faster and easier to perform.
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