Abstract
The telomeric repeat amplification protocol (TRAP) assay is a highly sensitive PCR based assay and is an important tool for understanding the role of telomerase in cancer. This assay measures an enzymatic activity where the amount of target is dependent upon the activity of the enzyme. This protocol consists of two steps: first, telomeric repeats are added to the substrate by the enzyme and second, the extended products will be amplified by Taq-DNA polymerase. The amplified TRAP assay products will be separated on 10% native PAGE and detected by SYBR Green I dye.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Greider, C.W., and Blackburn, E.H. 1989. A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis. Nature 337:331–337.
Kim, N.W., Piatyszek, M.A., Prowse, K.R., Harley, C.B., West, M.D., Ho, P.L., Coviello, G.M., Wright, W.E., Weinrich, S.L., and Shay, J.W. 1994. Specific association of human telomerase activity with immortal cells and cancer. Science 266:2011–2015.
Harley, C.B., and Villeponteau, B. 1995. Telomeres and telomerase in aging and cancer. Curr. Opin. Genet. Dev. 5 5:249–255.
Kilian, A., Bowtell, D.D., Abud, H.E., Hime, G.R., Venter, D.J., Keese, P.K., Duncan, E.L., Reddel, R.R., and Jefferson, R.A. 1997. Isolation of a candidate human telomerase catalytic subunit gene, which reveals complex splicing patterns in different cell types. Hum. Mol. Genet. 6:2011–2019.
Feng, J., 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., et al. 1995. The RNA component of human telomerase. Science 269:1236–1241.
Harrington, L., McPhail, T., Mar, V., Zhou, W., Oulton, R., Bass, M.B., Arruda, I., and Robinson, M.O. 1997. A mammalian telomerase-associated protein. Science 275:973–977.
Holt, S.E., Aisner, D.L., Baur, J., Tesmer, V.M., Dy, M., Ouellette, M., Trager, J.B., Morin, G.B., Toft, D.O., Shay, J.W., et al. 1999. Functional requirement of p23 and Hsp90 in telomerase complexes. Genes Dev. 13:817–826.
Nakano, K., Watney, E., and McDougall, J.K. 1998. Telomerase activity and expression of telomerase RNA component and telomerase catalytic subunit gene in cervical cancer. Am. J. Pathol. 153:857–864.
Boldrini, L., Faviana, P., Gisfredi, S., Zucconi, Y., Di Quirico, D., Donati, V., Berti, P., Spisni, R., Galleri, D., Materazzi, G., et al. 2002. Evaluation of telomerase in the development and progression of colon cancer. Int. J. Mol. Med. 10:589–592.
Park, T.W., Riethdorf, S., Riethdorf, L., Loning, T., and Janicke, F. 1999. Differential telomerase activity, expression of the telomerase catalytic sub-unit and telomerase-RNA in ovarian tumors. Int. J. Cancer 84:426–431.
Kanaya, T., Kyo, S., Takakura, M., Ito, H., Namiki, M., and Inoue, M. 1998. hTERT is a critical determinant of telomerase activity in renal-cell carcinoma. Int. J. Cancer 78:539–543.
Blackburn, E.H. 2001. Switching and signaling at the telomere. Cell 106:661–673.
Collins, K., and Mitchell, J.R. 2002. Telomerase in the human organism. Oncogene 21:564–579.
Blasco, M.A. 2005. Telomeres and human disease: ageing, cancer and beyond. Nat. Rev. Genet. 6:611–622.
Kim, N.W., and Wu, F. 1997. Advances in quantification and characterization of telomerase activity by the telomeric repeat amplification protocol (TRAP). Nucleic Acids Res. 25:2595–2597.
Jagadeesh, S., Kyo, S., and Banerjee, P.P. 2006. Genistein represses telomerase activity via both transcriptional and posttranslational mechanisms in human prostate cancer cells. Cancer Res. 66:2107–2115.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Banerjee, P.P., Jagadeesh, S. (2009). Non-Radioactive Assay Methods for the Assessment of Telomerase Activity and Telomere Length. In: Chellappan, S. (eds) Chromatin Protocols. Methods in Molecular Biology, vol 523. Humana Press. https://doi.org/10.1007/978-1-59745-190-1_25
Download citation
DOI: https://doi.org/10.1007/978-1-59745-190-1_25
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-58829-873-7
Online ISBN: 978-1-59745-190-1
eBook Packages: Springer Protocols