Citation: Zhang Jiayu, Zhou Xiaoyu, Zhou Man, Jia Hongxia. Detection of Telomerase Activity Based on Signal Amplification of Hybridization Chain Reaction Combining with Magnetic Separation[J]. Acta Chimica Sinica, ;2016, 74(6): 513-517. doi: 10.6023/A16030136 shu

Detection of Telomerase Activity Based on Signal Amplification of Hybridization Chain Reaction Combining with Magnetic Separation

1.
College of Chemistry and Environmental Science, Hebei University, Baoding 071002, China

Corresponding author: Jia Hongxia, jia123renren@126.com
Received Date: 18 March 2016

Fund Project: the Specialized Research Fund for the Doctoral Program of Higher Education of China 20121301120006the National Natural Science Foundation of China 21405032

Figures(5)

Telomerase is a ribonucleoprotein complex that is usually activated in the cancer cells and is closely related to telomere maintenance and immortalization of cancer cells. Telomerase activity detection is important for early diagnosis of human cancers as well as the screening of telomerase-target anti-cancer drugs. A new simple and fast method to detect the telomerase activity has been developed based on hybridization chain reaction (HCR) and magnetic separation. In the assay of experiment, the biotin-labeled telomerase substrate is elongated by telomerase generating a special DNA with repeated sequences-(ggttag)_n at their terminals. These telomerase elongated products are fixedly connected with streptavidin-coated magnetic beads through the specific combination of streptavidin with biotin. At the same time, other cell extracts are removed by magnetic separation. A specific DNA probe I is designed as the initiator of HCR. 3'-Terminus of DNA probe I is complementary with three repeated sequences of telomerase elongated product. So, DNA probe I could be fixed on magnetic bead through hybridization. 5'-Terminus of DNA probe I is in charge of triggering HCR with DNA probe II and probe III. DNA probe II and probe III are modified with fluorophores. So, the HCR amplification results can be easily detected by fluorescence. All of excessive DNA probes can be removed by magnetic separation. Under the optimal conditions, telomerase activity in 1.0×10⁵ Hela cells has been obviously detected. Because no enzyme involves in the signal amplification process of HCR, our proposed method can effectively avoid the interference of nonspecific amplification which usually exists in the polymerase amplification processes and increase the accuracy of the test results. Furthermore, enzyme-free signal amplification can effectively avoid the potential interference of telomerase inhibitor to the enzyme activity and improve the reliability of screening of telomerase inhibitors.
- telomerase activity,
- hybridization chain reaction,
- enzyme-free signal amplification,
- fluorescence detection,
- magnetic separation
1. [1]
  Cohen, S. B.; Graham, M. E.; Lovrecz, G. O.; Bache, N.; Robinson, P. J.; Reddel, R. R. Science 2007, 315, 1850. doi: 10.1126/science.1138596
2. [2]
  Feng, J. L.; Funk, W. D.; Wang, S. S.; Weinrich, S. L.; Avilion, A. A.; Chiu, C. P.; Adams, R. R.; Chang, E.; Allsopp, R. C.; Yu, J. H.; Le, S. Y.; West, M. D.; Harley, C. B.; Andrews, W. H.; Greider, C. W.; Villeponteau, B. Science 1995, 269, 1236. doi: 10.1126/science.7544491
3. [3]
  Liu, Y.; Chen, X. M.; Zhang, L. Q.; Sun, D. D.; Zhou, Y. H.; Chen, L. M.; Liu, J. Acta Chim. Sinica 2014, 72, 473. doi: 10.6023/A13101092
4. [4]
  Kim, N. W.; Piatyszek, M. A.; Prowse, K. R.; Harley, C. B.; West, M. D.; Ho, W. P. L. C.; Coviello, G. M.; Wright, W. E.; Weinrich, S. L.; Shay, J. W. Science 1994, 266, 2011. doi: 10.1126/science.7605428
5. [5]
  Kim, N. W.; Wu, F. Nucleic Acids Res. 1997, 25, 2595. doi: 10.1093/nar/25.13.2595
6. [6]
  Szatmari, I.; Tőkés, S.; Dunn, C. B.; Bardos, T. J.; Aradi, J. Anal. Biochem. 2000, 282, 80. doi: 10.1006/abio.2000.4589
7. [7]
  Szatmari, I.; Aradi, J. Nucleic Acids Res. 2001, 29, e3. doi: 10.1093/nar/29.2.e3
8. [8]
  Wege, H.; Chui, M. S.; Le, H. T.; Tran, J. M.; Zern, M. A. Nucleic Acids Res. 2003, 31, e3.
9. [9]
  Xiao, Y.; Dane, K. Y.; Uzawa, T.; Csordas, A.; Qian, J. R.; Soh, H. T.; Daugherty, P. S.; Lagally, E. T.; Heeger, A. J.; Plaxco, K. W. J. Am. Chem. Soc. 2010, 132, 15399. doi: 10.1021/ja107085n
10. [10]
  Huang, Y. P.; Kong, D. M.; Zhang, X. B.; Shen, H. X.; Mi, H. F. Acta Chim. Sinica 2004, 62, 274.
11. [11]
  Van Ness, J.; Van Ness, L. K.; Galas, D. J. Proc. Natl. Acad. Sci. U. S. A. 2003, 100, 4504. doi: 10.1073/pnas.0730811100
12. [12]
  Zhang, Y.; Wang, L. J.; Zhang, C. Y. Chem. Commun. 2014, 50, 1909. doi: 10.1039/c3cc48518h
13. [13]
  Tian, L. L.; Weizmann, Y. J. Am. Chem. Soc. 2013, 135, 1661. doi: 10.1021/ja309198j
14. [14]
  Ding, C. F.; Li, X. L.; Ge, Y.; Zhang, S. S. Anal. Chem. 2010, 82, 2850. doi: 10.1021/ac902818w
15. [15]
  Zhao, Y. X.; Qi, L.; Chen, F.; Zhao, Y.; Fan, C. H. Biosens. Bioelectron. 2013, 41, 764. doi: 10.1016/j.bios.2012.10.009
16. [16]
  Dirks, R. M.; Pierce, N. A. Proc. Natl. Acad. Sci. U. S. A. 2004, 101, 15275. doi: 10.1073/pnas.0407024101
17. [17]
  Niu, S. Y.; Jiang, Y.; Zhang, S. S. Chem. Commun. 2010, 46, 3089. doi: 10.1039/c000166j
18. [18]
  Chen, Y.; Xu, J.; Su, J.; Xiang, Y.; Yuan, R.; Chai, Y. Q. Anal. Chem. 2012, 84, 7750. doi: 10.1021/ac3012285
19. [19]
  Ge, Z. L.; Lin, M. H.; Wang, P.; Pei, H.; Yan, J.; Shi, J. Y.; Huang, Q.; He, D. N.; Fan, C. H.; Zuo, X. L. Anal. Chem. 2014, 86, 2124. doi: 10.1021/ac4037262
20. [20]
  Yang, L.; Liu, C. H.; Ren, W.; Li, Z. P. ACS Appl. Mater. Interfaces 2012, 4, 6450. doi: 10.1021/am302268t
21. [21]
  Zhang, B.; Liu, B. Q.; Tang, D. P.; Niessner, R.; Chen, G. N.; Knopp, D. Anal. Chem. 2012, 84, 5392. doi: 10.1021/ac3009065
22. [22]
  Wang, W. J.; Li, J. J.; Rui, K.; Gai, P. P.; Zhang, J. R.; Zhu, J. J. Anal. Chem. 2015, 87, 3019. doi: 10.1021/ac504652e
23. [23]
  Eskiocak, U.; Ozkan-Ariksoysal, D.; Ozsoz, M.; Öktem, H. A. Anal. Chem. 2007, 79, 8807. doi: 10.1021/ac071014r
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