Kar, Parmita ; Chawla, Himika ; Saha, Soma ; Tandon, Nikhil ; Goswami, Ravinder (2016) Identification of reference housekeeping-genes for mRNA expression studies in patients with type 1 diabetes Molecular and Cellular Biochemistry, 417 (1). pp. 49-56. ISSN 0300-8177
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Official URL: https://link.springer.com/article/10.1007/s11010-0...
Related URL: http://dx.doi.org/10.1007/s11010-016-2712-3
Abstract
Selection of appropriate housekeeping-genes as reference is important in mRNA expression-related experiments. It is more important in diabetes since hyperglycemia per se can influence expression of housekeeping-genes. RNA expression of Glyceraldehyde-3-phosphate-dehydrogenase, β-actin and 18S-ribosomal-RNA, Hypoxanthine-phosphoribosyl-transferase (HPRT), Tyrosine-3-monooxygenase/tryptophan (YHWAZ), β2-microglobin (β2M), TATA-binding-protein (TBP) and Ubiquitin C and cytochrome1 (CYC1) assessed in circulating-lymphocytes-(PBMC) of patients with type-1-diabetes and healthy controls. The stability (‘M’ value <1.02) and number of housekeeping-genes required for normalization in qRT-PCR were determined by ‘ge-norm software’. Vitamin-D-receptor (VDR) was used as a target gene. All the nine genes tested had sufficient ‘M’ value in diabetes and healthy controls. However, housekeeping-genes indicated a relatively higher stability of expression in healthy controls in comparison to diabetes. Use of single housekeeping-genes brought gross variation in the calculation of VDR-mRNA copies. The ge-norm software suggested geometric mean of five housekeeping-genes for ideal normalization in diabetes (CYC1, β-actin, YHWAZ, HPR and β2M) and only three in controls (CYC1, β-actin, and TBP). HbA1c did not correlate with expression of any of the nine housekeeping-genes. Thus, geometric mean of CYC1, β-actin, YHWAZ, HPRT and β2M needs to be used for ideal normalization of mRNA in type-1-diabetes. Similar studies are required in other population.
Item Type: | Article |
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Source: | Copyright of this article belongs to Springer-Verlag. |
Keywords: | Type 1 Diabetes; Housekeeping-genes; Gene Expression; Vitamin D Receptor |
ID Code: | 106876 |
Deposited On: | 27 Jun 2017 16:59 |
Last Modified: | 27 Jun 2017 16:59 |
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