Abstract

HOUSEKEEPING GENE AND ITS INTERNAL CONTROL

Kanika Adhikari*, Harshita Singh, Devanshu Sharma, Tusharika Arora and Vinod Kumar Gupta

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Abstract

Housekeeping genes are essential for maintaining the fundamental cellular processes required for cell survival and homeostasis. These genes are constitutively expressed and support core functions such as energy production, protein synthesis, structural maintenance, and DNA replication. They encode vital proteins like ribosomal components, actin, and chaperones involved in glycolysis, oxidative phosphorylation, and the citric acid cycle. In gene expression studies, housekeeping genes serve as internal controls in techniques like RT-PCR, enabling normalization of target gene expression while accounting for variations in sample quality, size, and experimental conditions. By minimizing technical variability, they ensure accurate comparisons across samples and verify that observed changes are not due to artifacts. Common examples include GAPDH, ACTB, and β2-microglobulin, which generally show stable expression across different cell types and conditions. However, the selection of suitable housekeeping genes requires careful consideration. Ideal reference genes should demonstrate consistent expression across tissues, remain unaffected by experimental treatments, and exhibit stability in the specific species or cell type under investigation. It's often recommended to use multiple housekeeping genes for accurate normalization and to avoid selecting genes involved in the pathway under study. Despite their widespread use, housekeeping genes can be influenced by the same variables that affect target genes, including disease states and treatment interventions. Experimental factors like RNA integrity and reverse transcription efficiency can also impact their quantification. Therefore, the assumption of uniform expression may not always hold true, highlighting the importance of validating housekeeping genes for each specific experimental context.

Keywords: Housekeeping, DNA Replication, GAPDH, RT-PCR, ACTB.