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Ovarian Cancer pp 171–186Cite as

Assessment of mRNA Splice Variants by qRT-PCR

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1049))

Abstract

Alternative splicing is an essential process for the generation of protein diversity. The physiological role, cellular localization, and abundance of splice variant products compared to the wild-type protein may be completely different. This is illustrated by the five splice variants of the antiapoptotic protein survivin that are more abundant in cancerous cells compared with normal tissues. Interestingly, some survivin splice variants have been associated with drug resistance. Herein, we describe a SYBR green I-based real-time PCR method to assess the messenger RNA levels of the human survivin splice variants in taxane-sensitive versus taxane-resistant ovarian cancer cells and in human ovarian cancer samples. Furthermore, in this chapter, we describe the quantification of survivin splice variants by real-time quantitative PCR (qPCR) after in vitro and in vivo small interference RNA (siRNA)-mediated silencing of survivin splice variants.

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Acknowledgments

We would like to thank Surangani Dharmawardhane Flanagan, Ph.D., for critical reading of the manuscript. This work was funded, in part, by the University of Puerto Rico Comprehensive Cancer Center seed funds to P.E.V.M. and a scholarship to I.M.E.V. from the NIH, Minority Biomedical Research Support-Research Initiative for Scientific Enhancement (MBRS-RISE) Program (R25-GM061838).

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Authors and Affiliations

  1. Department of Biochemistry, School of Medicine, University of Puerto Rico, San Juan, PR, USA

    Ileabett M. Echevarria Vargas

  2. Department of Biochemistry and Cancer Center, School of Medicine, University of Puerto Rico, San Juan, PR, USA

    Pablo E. Vivas-Mejía

Authors
  1. Ileabett M. Echevarria Vargas
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  2. Pablo E. Vivas-Mejía
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Editor information

Editors and Affiliations

  1. Group of Experimental Therapeutics, Federal State Institution N.N.Petrov Res Institute of Oncology, St. Petersburg, Russia

    Anastasia Malek

  2. Labs of Molecular Tumor Pathology & Func, Charité - Universitatsmedizin Berlin, Berlin, Germany

    Oleg Tchernitsa

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Vargas, I.M.E., Vivas-Mejía, P.E. (2013). Assessment of mRNA Splice Variants by qRT-PCR. In: Malek, A., Tchernitsa, O. (eds) Ovarian Cancer. Methods in Molecular Biology, vol 1049. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-547-7_13

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  • DOI: https://doi.org/10.1007/978-1-62703-547-7_13

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-546-0

  • Online ISBN: 978-1-62703-547-7

  • eBook Packages: Springer Protocols

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