Identification of housekeeping genes for microRNA expression analysis in kidney tissues of Pkd1 deficient mouse models
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 fiv...
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Published in | Scientific reports Vol. 10; no. 1; p. 231 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
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Nature Publishing Group UK
14.01.2020
Nature Publishing Group |
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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 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. |
---|---|
AbstractList | 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 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. 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 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. 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 :Nestin , CY), non-cystic controls (Pkd1 , NC), Pkd1-haploinsufficient (Pkd1 , HT), wild-type controls (Pkd1 , WT), severely cystic mice (Pkd1 , 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. 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 (Pkd1flox/flox:Nestincre, CY), non-cystic controls (Pkd1flox/flox, NC), Pkd1-haploinsufficient (Pkd1+/−, HT), wild-type controls (Pkd1+/+, WT), severely cystic mice (Pkd1V/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 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. |
ArticleNumber | 231 |
Author | Muñoz, J. J. Onuchic, L. F. Ormanji, M. S. Ferreira, F. M. Boim, M. A. Heilberg, I. P. Amaral, A. G. Anauate, A. C. Meca, R. |
Author_xml | – sequence: 1 givenname: J. J. orcidid: 0000-0002-1547-6357 surname: Muñoz fullname: Muñoz, J. J. organization: Nephrology Division, Department of Medicine, Universidade Federal de São Paulo – sequence: 2 givenname: A. C. surname: Anauate fullname: Anauate, A. C. organization: Nephrology Division, Department of Medicine, Universidade Federal de São Paulo – sequence: 3 givenname: A. G. surname: Amaral fullname: Amaral, A. G. organization: Divisions of Molecular Medicine and Nephrology, University of São Paulo School of Medicine – sequence: 4 givenname: F. M. surname: Ferreira fullname: Ferreira, F. M. organization: Laboratory of Immunology, Heart Institute, University of São Paulo School of Medicine – sequence: 5 givenname: R. surname: Meca fullname: Meca, R. organization: Nephrology Division, Department of Medicine, Universidade Federal de São Paulo – sequence: 6 givenname: M. S. orcidid: 0000-0003-1324-5800 surname: Ormanji fullname: Ormanji, M. S. organization: Nephrology Division, Department of Medicine, Universidade Federal de São Paulo – sequence: 7 givenname: M. A. surname: Boim fullname: Boim, M. A. organization: Nephrology Division, Department of Medicine, Universidade Federal de São Paulo – sequence: 8 givenname: L. F. surname: Onuchic fullname: Onuchic, L. F. organization: Divisions of Molecular Medicine and Nephrology, University of São Paulo School of Medicine – sequence: 9 givenname: I. P. surname: Heilberg fullname: Heilberg, I. P. email: ita.heilberg@gmail.com organization: Nephrology Division, Department of Medicine, Universidade Federal de São Paulo |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31937827$$D View this record in MEDLINE/PubMed |
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CitedBy_id | crossref_primary_10_1038_s41598_022_19548_z crossref_primary_10_3390_molecules29040748 crossref_primary_10_1038_s41598_021_99366_x crossref_primary_10_3390_ijms23147676 crossref_primary_10_1016_j_omtm_2023_101116 |
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Snippet | 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... Abstract Polycystic kidney disease is a complex clinical entity which comprises a group of genetic diseases that leads to renal cyst development. We evaluated... |
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SubjectTerms | 38 631/208/199 631/337/384/331 64 64/60 692/4022/1585/1589 Animal models Animals Gene Expression Profiling Genes, Essential - genetics Haploinsufficiency Humanities and Social Sciences Kidney - metabolism Kidney diseases Kidneys Mice MicroRNAs MicroRNAs - genetics miRNA multidisciplinary Polycystic kidney Protein Kinase C - deficiency Protein Kinase C - genetics Rodents Science Science (multidisciplinary) |
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Title | Identification of housekeeping genes for microRNA expression analysis in kidney tissues of Pkd1 deficient mouse models |
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