β-glucuronidase use as a single internal control gene may confound analysis in FMR1 mRNA toxicity studies
| العنوان: | β-glucuronidase use as a single internal control gene may confound analysis in FMR1 mRNA toxicity studies |
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| المؤلفون: | Claudine M Kraan, Xin Li, Kim Cornish, David E. Godler, Quang M. Bui, Howard R. Slater |
| المصدر: | PLoS ONE PLoS ONE, Vol 13, Iss 2, p e0192151 (2018) |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, Physiology, SDHA, lcsh:Medicine, Artificial Gene Amplification and Extension, Toxicology, Pathology and Laboratory Medicine, Biochemistry, Polymerase Chain Reaction, Fragile X Mental Retardation Protein, Mathematical and Statistical Techniques, Medicine and Health Sciences, lcsh:Science, Glucuronidase, education.field_of_study, Multidisciplinary, Messenger RNA, food and beverages, Confounding Factors, Epidemiologic, Body Fluids, Nucleic acids, Real-time polymerase chain reaction, Blood, Physical Sciences, Regression Analysis, RNA extraction, Anatomy, Statistics (Mathematics), Research Article, congenital, hereditary, and neonatal diseases and abnormalities, Nucleic acid synthesis, Population, Biology, Linear Regression Analysis, Real-Time Polymerase Chain Reaction, Research and Analysis Methods, Andrology, 03 medical and health sciences, Extraction techniques, Humans, RNA, Messenger, Chemical synthesis, Statistical Methods, RNA synthesis, education, Molecular Biology Techniques, Gene, Molecular Biology, Biology and life sciences, Toxicity, lcsh:R, fungi, RNA, Reverse Transcriptase-Polymerase Chain Reaction, FMR1, nervous system diseases, Biosynthetic techniques, 030104 developmental biology, lcsh:Q, Mathematics |
| الوصف: | Relationships between Fragile X Mental Retardation 1 (FMR1) mRNA levels in blood and intragenic FMR1 CGG triplet expansions support the pathogenic role of RNA gain of function toxicity in premutation (PM: 55-199 CGGs) related disorders. Real-time PCR (RT-PCR) studies reporting these findings normalised FMR1 mRNA level to a single internal control gene called β-glucuronidase (GUS). This study evaluated FMR1 mRNA-CGG correlations in 33 PM and 33 age- and IQ-matched control females using three normalisation strategies in peripheral blood mononuclear cells (PBMCs): (i) GUS as a single internal control; (ii) the mean of GUS, Eukaryotic Translation Initiation Factor 4A2 (EIF4A2) and succinate dehydrogenase complex flavoprotein subunit A (SDHA); and (iii) the mean of EIF4A2 and SDHA (with no contribution from GUS). GUS mRNA levels normalised to the mean of EIF4A2 and SDHA mRNA levels and EIF4A2/SDHA ratio were also evaluated. FMR1mRNA level normalised to the mean of EIF4A2 and SDHA mRNA levels, with no contribution from GUS, showed the most significant correlation with CGG size and the greatest difference between PM and control groups (p = 10-11). Only 15% of FMR1 mRNA PM results exceeded the maximum control value when normalised to GUS, compared with over 42% when normalised to the mean of EIF4A2 and SDHA mRNA levels. Neither GUS mRNA level normalised to the mean RNA levels of EIF4A2 and SDHA, nor to the EIF4A2/SDHA ratio were correlated with CGG size. However, greater variability in GUS mRNA levels were observed for both PM and control females across the full range of CGG repeat as compared to the EIF4A2/SDHA ratio. In conclusion, normalisation with multiple control genes, excluding GUS, can improve assessment of the biological significance of FMR1 mRNA-CGG size relationships. |
| تدمد: | 1932-6203 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::f5b7d6601b96c6c255de8cdd51690dba https://pubmed.ncbi.nlm.nih.gov/29474364 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....f5b7d6601b96c6c255de8cdd51690dba |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19326203 |
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