A single cell sensitive RT-PCR for the study of gene expression in mouse preimplantation development

Vittorio Sebastiano, Luca Gentile, Valeria Merico, Manuela Monti, Maurizio Zuccotti, Silvia Garagna, Carlo Alberto Redi

Research output: Contribution to journalArticle

Abstract

The study of gene expression during preimpkntation embryonic development is useful to understand which are the molecular mechanisms involved in the progressive passage from a maternal to a zygotic regulation of gene expression. It is our laboratory main concern to develop techniques that would allow a more precise characterisation of the timing of transcription and amount of transcripts of genes that are well known to play an important role in the early events of preimplantation development. In this study we illustrate the use of a very sensitive protocol of semiquantitative single-cell RT-PCR that we have developed. This technique allows the simultaneous analysis of up to eight genes on a single cell. Here we show the experiments performed to validate this novel assay. As an example we have analysed the quantitative expression of Hprt, Bax, Bex-1, Cpt-1, pt-2, Oct-4, Tsix and Xist on single blastocysts.

Original languageEnglish
Pages (from-to)117-126
Number of pages10
JournalRendiconti Lincei
Volume14
Issue number2
Publication statusPublished - 2003

Keywords

  • Gene expression
  • Preimplantation development
  • Retrotranscriptase-polymerase Chain Reaction

ASJC Scopus subject areas

  • Engineering(all)

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  • Cite this

    Sebastiano, V., Gentile, L., Merico, V., Monti, M., Zuccotti, M., Garagna, S., & Redi, C. A. (2003). A single cell sensitive RT-PCR for the study of gene expression in mouse preimplantation development. Rendiconti Lincei, 14(2), 117-126.