Selection of Candidate Housekeeping Genes for Normalization in Human Postmortem Brain Samples
Open accessAbstract
The most frequently used technique to study the expression profile of genes involved in common neurological disorders is quantitative real-time RT-PCR, which allows the indirect detection of very low amounts of selected mRNAs in tissue samples. Expression analysis by RT-qPCR requires an appropriate normalization to the expression level of genes characterized by a stable, constitutive transcription. However, the identification of a gene transcribed at a very stable level is difficult if not impossible, since significant fluctuations of the level of mRNA synthesis often accompanies changes of cell behavior. The aim of this study is to identify the most stable genes in postmortem human brain samples of patients affected by Alzheimer’s disease (AD) suitable as reference genes. The experiments analyzed 12 commonly used reference genes in brain samples from eight individuals with AD and seven controls. After a careful analysis of the results calculated by geNorm and NormFinder algorithms, we found that CYC1 and EIF4A2 are the best reference genes. We remark on the importance of the determination of the best reference genes for each sample to be analyzed and suggest a practical combination of reference genes to be used in the analysis of human postmortem samples.
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EIFA4 and CYC1 were selected as internal controls for human RT-PCR based upon their stability in postmortem brain of AD patients.
“EIFA4 and CYC1 were selected as internal controls for human RT-PCR based upon their stability in postmortem brain of AD patients54.”
From Tau promotes neurodegeneration through global chromatin relaxation (2014) · cites this paper as [54]
The Results identify CYC1 as a stable reference gene in postmortem brain samples from AD patients and controls. The Discussion recommends CYC1 together with EIF4A2 for RT-qPCR normalization, but the claim names EIFA4 instead of EIF4A2. (LLM, conf=0.88)
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- Authors
- Penna, I., Vella, S., Gigoni, A., Russo, C., Cancedda, R., & Pagano, A.
- Affiliations
- University of Genoa (IT)
- University of Molise (IT)
- Corresponding author
- Pagano, Aldo
- Venue
- International Journal of Molecular Sciences
- Publisher
- MDPI AG
- Published
- 2011-08-26
- Volume
- 12
- Issue
- 9
- Pages
- 5461-5470
- ISSN
- 1422-0067 (ISSN-L)
- PMC
- PMC3189726
- OpenAlex
- W2082658803
- Cited by
- 79
- References
- 0
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 3 · 2023: 2 · 2022: 4 · 2021: 9 · 2020: 7 · 2019: 5 · 2018: 6 · 2017: 6 · 2016: 8 · 2015: 3 · 2014: 5 · 2013: 8 · 2012: 8
- FWCI
- 2.72
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- International Journal of Molecular Sciences (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- CINECA IRIS Institutial Research Information System (University of Genoa) (submittedVersion)
- Archivio Istituzionale della Ricerca - Università degli Studi del Molise (submittedVersion)
- Funders
- Ministero dell’Istruzione, dell’Università e della Ricerca (2007945BZN)
- Associazione Italiana per la Ricerca sul Cancro (IG9378)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Molecular Biology Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene expression and cancer classification
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- reference genes, housekeeping genes, gene expression normalization, postmortem brain, Alzheimer's disease, cytochrome c1, EIF4A2, geNorm, NormFinder, RT-qPCR, genetic stability
- MeSH terms
- Aged, Aged, 80 and over, Algorithms, Alzheimer Disease, Brain, Cytochromes c1, Female, Humans, Male, Middle Aged, Postmortem Changes, Reference Standards, Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, Eukaryotic Initiation Factor-4A
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