Optimal endogenous controls for microRNA analysis of visceral adipose tissue in the NAFLD mouse model
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00179906%3A_____%2F25%3A10495128" target="_blank" >RIV/00179906:_____/25:10495128 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11150/25:10495128
Result on the web
<a href="https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=52wjYkQ1W7" target="_blank" >https://verso.is.cuni.cz/pub/verso.fpl?fname=obd_publikace_handle&handle=52wjYkQ1W7</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1007/s12038-025-00492-9" target="_blank" >10.1007/s12038-025-00492-9</a>
Alternative languages
Result language
angličtina
Original language name
Optimal endogenous controls for microRNA analysis of visceral adipose tissue in the NAFLD mouse model
Original language description
The selection of proper reference genes and materials is critical in the design of PCR experiments, especially for differential expression studies. In this study, we propose a method to identify robust endogenous control miRNAs in the visceral adipose tissue of C57BL/6J mice with non-alcoholic fatty liver disease induced by alternating Western and control diets. This study outlines a comprehensive methodology for the analysis of microRNA endogenous controls using microfluidic cards in conjunction with miRNA profiling through small RNA sequencing and subsequent validation by quantitative PCR and the RefFinder algorithm. Criteria included were fold change, p-value, reads per million, and gene stability assessment. A set of six putative endogenous microRNAs was identified (miR-331-3p, let-7a-5p, miR-1839-5p, miR-151a-5p, let-7d-5p, and let-7c-5p). Subsequent validation and analysis using the RefFinder algorithm assessed the stability of the selected genes, and a combination of the three most stable endogenous miRNA controls (miR-331-3p, let-7a-5p, and miR-1839-5p) exhibiting consistent expression patterns with minimal variability was set. Given the absence of universal endogenous controls, individual evaluation of normalizers for each experiment is imperative for accurate miRNA expression measurements. This approach, which combines multiple techniques and assessments, provides a reliable strategy for identifying and validating endogenous controls in miRNA studies.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
30105 - Physiology (including cytology)
Result continuities
Project
<a href="/en/project/NU21-07-00550" target="_blank" >NU21-07-00550: Ketogenic diet as a treatment for inborn defects of ATP synthase</a><br>
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2025
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Journal of Biosciences
ISSN
0250-5991
e-ISSN
0973-7138
Volume of the periodical
50
Issue of the periodical within the volume
1
Country of publishing house
IN - INDIA
Number of pages
14
Pages from-to
11
UT code for WoS article
001425887900002
EID of the result in the Scopus database
2-s2.0-85218468792