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...
Saved in:
Published in | Scientific reports Vol. 10; no. 1; p. 231 |
---|---|
Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
London
Nature Publishing Group UK
14.01.2020
Nature Publishing Group |
Subjects | |
Online Access | Get full text |
Cover
Loading…
Summary: | 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. |
---|---|
Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/s41598-019-57112-4 |