Abstract
The TP53 gene expresses at least nine different mRNA variants (p53 isoform mRNAs), including the one encoding the canonical p53 tumor suppressor protein. We have developed scientific tools to specifically detect and quantify p53 isoform expression at mRNA level by nested RT-PCR (reverse transcription-polymerase chain reaction) and quantitative real-time RT-PCR (RT-qPCR using the TaqMan® chemistry). Here, we describe these two methods, while highlighting essential points with regard to the analysis of p53 isoform mRNA expression.
Marie P. Khoury and Virginie Marcel have contributed equally to this work.
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Acknowledgments
This work was supported by Cancer Research UK (C8/A6613). M.K., K.F., A.D., D.L., and J.C.B. are supported by Cancer Research UK and V.M. is supported by Breast Cancer Campaign.
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Khoury, M.P., Marcel, V., Fernandes, K., Diot, A., Lane, D.P., Bourdon, JC. (2013). Detecting and Quantifying p53 Isoforms at mRNA Level in Cell Lines and Tissues. In: Deb, S., Deb, S. (eds) p53 Protocols. Methods in Molecular Biology, vol 962. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-236-0_1
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DOI: https://doi.org/10.1007/978-1-62703-236-0_1
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