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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Identification of housekeeping genes for microRNA expression analysis in kidney tissues of Pkd1 deficient mouse models

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Author(s):
Munoz, J. J. [1] ; Anauate, A. C. [1] ; Amaral, A. G. [2, 3] ; Ferreira, F. M. [4] ; Meca, R. [1] ; Ormanji, M. S. [1] ; Boim, M. A. [1] ; Onuchic, L. F. [2, 3] ; Heilberg, I. P. [1]
Total Authors: 9
Affiliation:
[1] Univ Fed Sao Paulo, Dept Med, Nephrol Div, Sao Paulo - Brazil
[2] Univ Sao Paulo, Div Mol Med, Sch Med, Sao Paulo - Brazil
[3] Univ Sao Paulo, Div Nephrol, Sch Med, Sao Paulo - Brazil
[4] Univ Sao Paulo, Heart Inst, Lab Immunol, Sch Med, Sao Paulo - Brazil
Total Affiliations: 4
Document type: Journal article
Source: SCIENTIFIC REPORTS; v. 10, n. 1 JAN 14 2020.
Web of Science Citations: 0
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(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 Delta Ct 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. (AU)

FAPESP's process: 15/17152-3 - Smoking Effects on the Renal and Cardiac Phenotypes in Cystic Mice due to Pkd1 Inactivation
Grantee:Luiz Fernando Onuchic
Support Opportunities: Regular Research Grants
FAPESP's process: 11/21593-4 - Caffeine effects on polycystic kidney disease development and progression, using a murine model Pkd1 cond/cond: Balcre
Grantee:Ita Pfeferman Heilberg
Support Opportunities: Regular Research Grants
FAPESP's process: 18/09135-0 - Evaluation of fibrosis factors and microRNAs in the progression of the polycystic renal disease in a PKD1 deficient murine model
Grantee:Juan José Augusto Moyano Muñoz
Support Opportunities: Scholarships in Brazil - Post-Doctoral
FAPESP's process: 15/23345-9 - MicroRNAs, extracellular vesicles and stem cells: physiology, pathophysiological role and therapeutic potential in renal diseases
Grantee:Mirian Aparecida Boim
Support Opportunities: Research Projects - Thematic Grants