First Author | Muñoz JJ | Year | 2020 |
Journal | Sci Rep | Volume | 10 |
Issue | 1 | Pages | 231 |
PubMed ID | 31937827 | Mgi Jnum | J:292914 |
Mgi Id | MGI:6407228 | Doi | 10.1038/s41598-019-57112-4 |
Citation | Munoz JJ, et al. (2020) Identification of housekeeping genes for microRNA expression analysis in kidney tissues of Pkd1 deficient mouse models. Sci Rep 10(1):231 |
abstractText | 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(flox/flox):Nestin(cre), CY), non-cystic controls (Pkd1(flox/flox), NC), Pkd1-haploinsufficient (Pkd1(+/-), HT), wild-type controls (Pkd1(+/+), WT), severely cystic mice (Pkd1(V/V), 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 DeltaCt 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. |