Selection and Expression Stability Analysis of Housekeeping Genes for Real-Time PCR Normalization in Rhesus Macaque...

Area of Science:

• Primate research
• Molecular biology
• Genomics

Background:

• Rhesus macaques (Macaca mulatta) are vital models in translational research.
• Limited molecular tools hinder physiological status monitoring in captive macaques.
• Validated housekeeping genes (HKGs) are lacking for mRNA expression analysis in rhesus macaque blood.

Purpose of the Study:

• To identify and validate stable housekeeping genes (HKGs) for gene expression studies in Macaca mulatta.
• To establish reliable reference genes for peripheral blood mRNA analysis in this species.
• To improve health monitoring and population management of captive rhesus macaques.

Main Methods:

• Candidate HKG expression stability was assessed using real-time PCR and RefFinder analysis.
• Peripheral blood samples from healthy adult male rhesus macaques were utilized.
• A multistage selection process identified the most stable genes from an initial panel of 29 candidates.

Main Results:

• Seven genes, including AHSA1, B4GALT3, CYB5R1, HPCAL1, PSMD6, SSR2, and VPS72, were evaluated for expression stability.
• B4GALT3 and PSMD6 demonstrated the highest expression stability among the tested candidates.
• 22 out of 29 candidate genes were excluded after rigorous bioinformatic and experimental validation.

Conclusions:

• This study provides an experimentally validated panel of HKGs for Macaca mulatta.
• B4GALT3 and PSMD6 are recommended as reliable reference genes for gene expression studies in rhesus macaque peripheral blood.
• The findings will enhance the accuracy of gene expression analysis, aiding in macaque health monitoring and research.