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Identification of housekeeping genes for microRNA expression analysis in kidney tissues of Pkd1 deficient mouse models.
- Source :
-
Scientific reports [Sci Rep] 2020 Jan 14; Vol. 10 (1), pp. 231. Date of Electronic Publication: 2020 Jan 14. - Publication Year :
- 2020
-
Abstract
- Polycystic kidney disease is a complex clinical entity which comprises a group of genetic diseases that leads to renal cyst development. We evaluated the most suitable housekeeping genes for microRNA expression by RT-qPCR analyses of kidney tissues in Pkd1-deficient mouse models from a panel of five candidates genes (miR-20a, miR-25, miR-26a, miR-191 and U6) and 3 target genes (miR-17, miR-21 and let-7a) using samples from kidneys of cystic mice (Pkd1 <superscript>flox/flox</superscript> :Nestin <superscript>cre</superscript> , CY), non-cystic controls (Pkd1 <superscript>flox/flox</superscript> , NC), Pkd1-haploinsufficient (Pkd1 <superscript>+/-</superscript> , HT), wild-type controls (Pkd1 <superscript>+/+</superscript> , WT), severely cystic mice (Pkd1 <superscript>V/V</superscript> , SC), wild-type controls (CO). The stability of the candidate genes was investigated using NormFinder, GeNorm, BestKeeper, DataAssist, and RefFinder software packages and the comparative ΔCt method. The analyses identified miR-26a as the most stable housekeeping gene for all kidney samples, miR-20a for CY and NC, miR-20a and miR-26a for HT and WT, and miR-25 and miR-26a for SC and CO. Expression of miR-21 was upregulated in SC compared to CO and trends of miR-21 upregulation and let-7a downregulation in CY and HT compared to its control kidneys, when normalized by different combinations of miR-20a, miR-25 and miR-26a. Our findings established miR-20a, miR-25, and miR-26a as the best housekeeping genes for miRNA expression analyses by RT-qPCR in kidney tissues of Pkd1-deficient mouse models.
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 10
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 31937827
- Full Text :
- https://doi.org/10.1038/s41598-019-57112-4