Protocol

p53 Protocols

Volume 962 of the series Methods in Molecular Biology pp 1-14

Date:

Detecting and Quantifying p53 Isoforms at mRNA Level in Cell Lines and Tissues

  • Marie P. KhouryAffiliated withCR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee
  • , Virginie MarcelAffiliated withCR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee
  • , Kenneth FernandesAffiliated withCR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee
  • , Alexandra DiotAffiliated withCR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee
  • , David P. LaneAffiliated withCR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee
  • , Jean-Christophe BourdonAffiliated withDepartment of Surgery and Molecular Oncology, Ninewells Hospital, University of Dundee Email author 

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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.

Key words

p53 isoforms Cancer Detection Quantification mRNA Splicing Nested PCR Quantitative real-time RT-PCR (TaqMan® chemistry) p53 tumor suppressor protein