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Screening for miRNA Expression Changes Using Quantitative PCR (Q-PCR)

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Cancer Epigenetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 863))

Abstract

In this section, we describe the use of Applied Biosystems TaqMan Array microRNA Card Set 3.0 to identify miRNA expression in a given RNA sample. This array set includes an array “A” and an array “B” which each have 384 wells that contain specific forward, reverse, and probe oligoinucleotides for measuring the expression of individual miRNAs during a Real-Time PCR (Q-PCR) reaction. Array “A” includes assays for profiling the comparatively higher expressed and better characterized miRNAs. Presently there are 1,048 mature miRNAs annotated in miRBase (release 16). The relatively small amount of miRNAs in comparison to protein-coding genes makes this format a viable option for measuring genome-wide miRNA expression changes. The Applied Biosystems TaqMan miRNA array set 3.0, which includes two separate arrays, can profile 754 miRNAs.

An erratum to this chapter is available at http://dx.doi.org/10.1007/978-1-61779-612-8_29

An erratum to this chapter can be found at http://dx.doi.org/10.1007/978-1-61779-612-8_29

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References

  1. Johnson, C. D., Esquela-Kerscher, A., Stefani, G., Byrom, M., Kelnar, K., Ovcharenko, D., Wilson, M., Wang, X., Shelton, J., Shingara, J., Chin, L., Brown, D., and Slack, F. J. (2007) The let-7 microRNA represses cell proliferation pathways in human cells, Cancer Res 67, 7713–7722.

    Article  PubMed  CAS  Google Scholar 

  2. Cimmino, A., Calin, G. A., Fabbri, M., Iorio, M. V., Ferracin, M., Shimizu, M., Wojcik, S. E., Aqeilan, R. I., Zupo, S., Dono, M., Rassenti, L., Alder, H., Volinia, S., Liu, C. G., Kipps, T. J., Negrini, M., and Croce, C. M. (2005) miR-15 and miR-16 induce apoptosis by targeting BCL2, Proc Natl Acad Sci U S A 102, 13944–13949.

    Article  PubMed  CAS  Google Scholar 

  3. Esau, C., Kang, X., Peralta, E., Hanson, E., Marcusson, E. G., Ravichandran, L. V., Sun, Y., Koo, S., Perera, R. J., Jain, R., Dean, N. M., Freier, S. M., Bennett, C. F., Lollo, B., and Griffey, R. (2004) MicroRNA-143 regulates adipocyte differentiation, J Biol Chem 279, 52361–52365.

    Article  PubMed  CAS  Google Scholar 

  4. Ma, L., Teruya-Feldstein, J., and Weinberg, R. A. (2007) Tumour invasion and metastasis initiated by microRNA-10b in breast cancer, Nature 449, 682–688.

    Article  PubMed  CAS  Google Scholar 

  5. Griffiths-Jones, S., Saini, H. K., van Dongen, S., and Enright, A. J. (2008) miRBase: tools for microRNA genomics, Nucleic Acids Res 36, D154–158.

    Article  PubMed  CAS  Google Scholar 

  6. Calin, G. A. and Croce, C. M. (2006) MicroRNA signatures in human cancers, Nat Rev Cancer 6, 857–866.

    Article  PubMed  CAS  Google Scholar 

  7. Suzuki, H., Yamamoto, E., Nojima, M., Kai, M., Yamano, H. O., Yoshikawa, K., Kimura, T., Kudo, T., Harada, E., Sugai, T., Takamaru, H., Niinuma, T., Maruyama, R., Yamamoto, H., Tokino, T., Imai, K., Toyota, M., and Shinomura, Y. Methylation-associated silencing of microRNA-34b/c in gastric cancer and its involvement in an epigenetic field defect, Carcinogenesis 31, 2066–2073.

    Google Scholar 

  8. Lujambio, A., Calin, G. A., Villanueva, A., Ropero, S., Sanchez-Cespedes, M., Blanco, D., Montuenga, L. M., Rossi, S., Nicoloso, M. S., Faller, W. J., Gallagher, W. M., Eccles, S. A., Croce, C. M., and Esteller, M. (2008) A microRNA DNA methylation signature for human cancer metastasis, Proc Natl Acad Sci U S A 105, 13556–13561.

    Article  PubMed  CAS  Google Scholar 

  9. Saito, Y., Suzuki, H., Tsugawa, H., Nakagawa, I., Matsuzaki, J., Kanai, Y., and Hibi, T. (2009) Chromatin remodeling at Alu repeats by epigenetic treatment activates silenced microRNA-512-5p with downregulation of Mcl-1 in human gastric cancer cells, Oncogene 28, 2738–2744.

    Article  PubMed  CAS  Google Scholar 

  10. Biosystems, A. Megaplex™ Pools For microRNA Expression Analysis Protocol (P/N: 4399721).

    Google Scholar 

  11. Biosystems, A. User Bulletin: Applied Biosystems Taqman Low Density Array.

    Google Scholar 

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Correspondence to Richard N. Armstrong .

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Armstrong, R.N., Colyer, H.A.A., Mills, K.I. (2012). Screening for miRNA Expression Changes Using Quantitative PCR (Q-PCR). In: Dumitrescu, R., Verma, M. (eds) Cancer Epigenetics. Methods in Molecular Biology, vol 863. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-612-8_18

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  • DOI: https://doi.org/10.1007/978-1-61779-612-8_18

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-611-1

  • Online ISBN: 978-1-61779-612-8

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