CAS OpenIR  > 中科院上海应用物理研究所2011-2019年
Highly sensitive detection of telomerase activity in tumor cells by cascade isothermal signal amplification based on three-way junction and base-stacking hybridization
Zhao, Yongxi; Qi, Lin; Chen, Feng; Zhao, Yue; Fan, Chunhai(樊春海)
2013
Source PublicationBIOSENSORS & BIOELECTRONICS
ISSN0956-5663
Volume41Pages:CONCATENATE(Sheet1!I149,-Sheet1!J149)
AbstractHerein, We report a simple and highly sensitive telomerase activity assay that integrates two consecutive isothermal signal amplification processes, namely, three-way junction triggered DNA-machine (3WJ-DNAM), and base-stacking hybridization assisted "biological circuit" DNA-machine (BSHBC-DNAM). In the presence of telomerase, the 3WJ are formed by the hybridization between the telomerase product and 3WJ-probes (3WJ-primer and 3WJ-template), which will initiate an autonomous 3WJ-DNAM by multiple processes of replication, nicking, and strand displacement, continuously generating short oligonucleotides as "triggers". These "triggers" will then provide additional stability for another two primers with a shared 5-bp complementary sequence at each 3'-end via base-stacking hybridization. And the BSHBC-DNAM are subsequently carried out by the strand-displacement induced circular utilization of "Trigger". Eventually, the single-stranded DNA (ssDNA) is generated in large quantities, and a significant fluorescence enhancement is observed due to the hybridization between the ssDNA and molecular beacons (MBs). In this way, per telomerase-mediated elongation event is efficiently and specifically converted into the greatly amplified fluorescence signals. This novel sensing strategy permits measurement of telomerase activity in cell extracts over the range of 3-5000 Hela cells, which is comparable or even superior to most previously reported methods. Using somatic and tumor cell lines, the selectivity and generality of the assay are investigated with satisfactory results. Furthermore, the inhibition effect of 3'-azido-3'-deoxythymidine (AZT) is also investigated, indicating its excellent performance in telomerase inhibitor screening. (C) 2012 Elsevier B.V. All rights reserved.
Indexed BySCI
Language英语
Funding Project应物所项目组
WOS IDWOS:000314191300114
Citation statistics
Cited Times:54[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.sinap.ac.cn/handle/331007/13766
Collection中科院上海应用物理研究所2011-2019年
Recommended Citation
GB/T 7714
Zhao, Yongxi,Qi, Lin,Chen, Feng,et al. Highly sensitive detection of telomerase activity in tumor cells by cascade isothermal signal amplification based on three-way junction and base-stacking hybridization[J]. BIOSENSORS & BIOELECTRONICS,2013,41:CONCATENATE(Sheet1!I149,-Sheet1!J149).
APA Zhao, Yongxi,Qi, Lin,Chen, Feng,Zhao, Yue,&Fan, Chunhai.(2013).Highly sensitive detection of telomerase activity in tumor cells by cascade isothermal signal amplification based on three-way junction and base-stacking hybridization.BIOSENSORS & BIOELECTRONICS,41,CONCATENATE(Sheet1!I149,-Sheet1!J149).
MLA Zhao, Yongxi,et al."Highly sensitive detection of telomerase activity in tumor cells by cascade isothermal signal amplification based on three-way junction and base-stacking hybridization".BIOSENSORS & BIOELECTRONICS 41(2013):CONCATENATE(Sheet1!I149,-Sheet1!J149).
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