Identification of suitable reference genes in the mouse placenta

Area of Science:

• Reproductive Biology
• Genomics
• Molecular Biology

Background:

• Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) is crucial for analyzing gene expression.
• Housekeeping genes are typically used as references, assuming stable expression.
• No prior validation exists for reference genes in mouse placenta studies.

Purpose of the Study:

• To empirically validate the stability of candidate housekeeping genes for RT-qPCR in mouse placenta.
• To identify the most reliable reference genes for accurate gene expression analysis in this tissue.

Main Methods:

• Analyzed expression stability of 32 candidate housekeeping genes in mouse placentas using NormFinder and BestKeeper software.
• Quantified cycle threshold (Ct) values via RT-qPCR in various experimental conditions and time points.

Main Results:

• Fifteen genes showed optimal Ct values; B2m, Polr2a, Ubc, and Ywhaz exhibited high stability.
• Polr2a demonstrated the highest expression stability overall.
• Polr2a combined with Ubc showed excellent stability in mouse placental tissue.

Conclusions:

• Polr2a, particularly with Ubc, is highly suitable as an endogenous reference for gene expression analysis in mouse placenta.
• Validation in human placenta confirms the translational relevance of these findings.