<i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic Differentiation
RT-qPCR is the gold standard and the most commonly used method for measuring gene expression. Selection of appropriate reference gene(s) for normalization is a crucial part of RT-qPCR experimental design, which allows accurate quantification and reliability of the results. Because there is no univer...
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MDPI AG
2022-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/8/4257 |
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| author | Nina Franko Lucija Ana Vrščaj Taja Zore Barbara Ostanek Janja Marc Jasna Lojk |
| author_facet | Nina Franko Lucija Ana Vrščaj Taja Zore Barbara Ostanek Janja Marc Jasna Lojk |
| author_sort | Nina Franko |
| collection | DOAJ |
| description | RT-qPCR is the gold standard and the most commonly used method for measuring gene expression. Selection of appropriate reference gene(s) for normalization is a crucial part of RT-qPCR experimental design, which allows accurate quantification and reliability of the results. Because there is no universal reference gene and even commonly used housekeeping genes’ expression can vary under certain conditions, careful selection of an appropriate internal control must be performed for each cell type or tissue and experimental design. The aim of this study was to identify the most stable reference genes during osteogenic differentiation of the human osteosarcoma cell lines MG-63, HOS, and SaOS-2 using the geNorm, NormFinder, and BestKeeper statistical algorithms. Our results show that <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i>, and <i>EF1A1</i> are the most stably expressed genes, while <i>ACTB</i>, and <i>18S rRNA</i> expressions are most variable. These data provide a basis for future RT-qPCR normalizations when studying gene expression during osteogenic differentiation, for example, in studies of osteoporosis and other bone diseases. |
| first_indexed | 2024-03-09T13:33:23Z |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T13:33:23Z |
| publishDate | 2022-04-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a0134eaf63e74fa1808107d5949369fa2023-11-30T21:15:17ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-04-01238425710.3390/ijms23084257<i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic DifferentiationNina Franko0Lucija Ana Vrščaj1Taja Zore2Barbara Ostanek3Janja Marc4Jasna Lojk5Department of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, SloveniaDepartment of Clinical Biochemistry, Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, SloveniaRT-qPCR is the gold standard and the most commonly used method for measuring gene expression. Selection of appropriate reference gene(s) for normalization is a crucial part of RT-qPCR experimental design, which allows accurate quantification and reliability of the results. Because there is no universal reference gene and even commonly used housekeeping genes’ expression can vary under certain conditions, careful selection of an appropriate internal control must be performed for each cell type or tissue and experimental design. The aim of this study was to identify the most stable reference genes during osteogenic differentiation of the human osteosarcoma cell lines MG-63, HOS, and SaOS-2 using the geNorm, NormFinder, and BestKeeper statistical algorithms. Our results show that <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i>, and <i>EF1A1</i> are the most stably expressed genes, while <i>ACTB</i>, and <i>18S rRNA</i> expressions are most variable. These data provide a basis for future RT-qPCR normalizations when studying gene expression during osteogenic differentiation, for example, in studies of osteoporosis and other bone diseases.https://www.mdpi.com/1422-0067/23/8/4257reference genegeNormNormFinderBestKeeperosteogenic differentiationgene expression |
| spellingShingle | Nina Franko Lucija Ana Vrščaj Taja Zore Barbara Ostanek Janja Marc Jasna Lojk <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic Differentiation International Journal of Molecular Sciences reference gene geNorm NormFinder BestKeeper osteogenic differentiation gene expression |
| title | <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic Differentiation |
| title_full | <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic Differentiation |
| title_fullStr | <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic Differentiation |
| title_full_unstemmed | <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic Differentiation |
| title_short | <i>TBP</i>, <i>PPIA</i>, <i>YWHAZ</i> and <i>EF1A1</i> Are the Most Stably Expressed Genes during Osteogenic Differentiation |
| title_sort | i tbp i i ppia i i ywhaz i and i ef1a1 i are the most stably expressed genes during osteogenic differentiation |
| topic | reference gene geNorm NormFinder BestKeeper osteogenic differentiation gene expression |
| url | https://www.mdpi.com/1422-0067/23/8/4257 |
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