Use of Chuk as an internal standard suitable for quantitative RT-PCR in mouse preimplantation embryos

Geppino Falco, Ilaria Stanghellini, Minoru S.H. Ko

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Analysis of gene expression changes during preimplantation development by quantitative reverse transcription-polymerase chain reaction (Q-PCR) requires appropriate internal standards. Ideally, such a gene should show a constant level of transcripts per embryo across all preimplantation stages from unfertilized eggs to blastocysts. By analysing the microarray-based gene expression profiles of preimplantation embryos, it was found that a conserved helix-loop-helix ubiquitous kinase gene (Chuk, also known as IκB kinase α, IKKα or IKK1) satisfied this criterion. To test the utility of this gene as an internal standard for Q-PCR, the expression levels of two known genes (Nalp5/Mater, Pou5f1/Oct3/Oct4) were normalized by Chuk and other housekeeping genes (Actb, Gapdh, Eef1a1, and H2afz) and demonstrated that the former was more consistent with the expression patterns obtained by a whole-mount in-situ hybridization than those reported previously with the latter. It is concluded that Chuk, unlike other commonly used normalization controls, is a reliable and suitable internal standard for measuring gene expression levels by Q-PCR in mouse oocytes and preimplantation embryos.

Original languageEnglish
Article number2376
Pages (from-to)394-403
Number of pages10
JournalReproductive BioMedicine Online
Volume13
Issue number3
DOIs
Publication statusPublished - 2006 Sep
Externally publishedYes

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Keywords

  • Gene expression
  • Internal standard
  • Mouse preimplantation
  • Normalization
  • Q-PCR

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynaecology
  • Developmental Biology

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