Identification of endogenous microRNA references in porcine serum for quantitative real-time PCR normalization

Bae, In-Seon; Seo, Kang-Seok; Kim, Sang Hoon

doi:10.1007/s11033-018-4242-4

Cited by 6 publications

(4 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…There is a diverse opinion on reference gene selection for PCR-based miRNA gene expression studies [7,32,33]. Different normalization methods are in use for the selection of reference gene for miRNA expression studies [5,34].…”

Section: Discussionmentioning

confidence: 99%

Mean Expression Value Normalized and Absolute Quantified miR-21 Found to be a Potential Diagnostic and Prognostic Marker for IgA Nephropathy

Kumar¹

2020

IJST

View full text Add to dashboard Cite

Background/objectives: miR-21 has been linked to renal fibrosis and miR-155 too has shown different expression in IgAN whose pathogenesis is partially understood. In this study, we explored the utility of miR-21 and miR-155 in IgA nephropathy. Methods: We recruited 40 IgA nephropathy (IgAN) patients and 15 healthy controls (HC). 2 ml blood sample was collected from each study participant. Mean expressed value normalization and absolute and relative quantification methods were used for quantitative polymerase chain reaction (PCR) experiment for miRNA expression. Logistic regression analysis was performed for diagnostic and prognostic value of miR-21. Findings: Mean expressed value normalization reduced the variation in groups and between groups without changing the mean value. miR-155 showed similar expression in IgAN and healthy controls. There was threefold increases in miR-21 in IgAN than the HC. miR-21 was able to differentiate the mesangial hypercellularity and tubular atrophy/interstitial fibrosis stages with sensitivity 84 and 70 percent, respectively. Applications: miR-155 can be used as a reference control gene for IgAN on quantitative PCR-based experiments. miR-21 can be used as a diagnostic and prognostic marker for IgAN.

show abstract

Section: Discussionmentioning

confidence: 99%

Mean Expression Value Normalized and Absolute Quantified miR-21 Found to be a Potential Diagnostic and Prognostic Marker for IgA Nephropathy

Kumar¹

2020

IJST

View full text Add to dashboard Cite

show abstract

“…We performed qRT-PCR in the Applied Biosystems ® QuantStudio™ 7 Flex Real-Time PCR System (Thermo Fisher, Singapore) using PowerUp™ SYBR™ Green Master Mix (Thermo Fisher, Vilnius, Lithuania). As previously studied, U6 and Hypoxanthine Phosphoribosyltransferase 1 ( HPRT1 ) were used as internal reference genes for miRNA and mRNA, respectively [ 67 , 68 ]; meanwhile, according to the previous miRNA and mRNA sequencing data, U6 and HPRT1 were stably expressed in AE and HE with similar expression levels. In summary, we selected the above two genes as internal reference genes.…”

Section: Methodsmentioning

confidence: 99%

Differential MicroRNA Expression in Porcine Endometrium Related to Spontaneous Embryo Loss during Early Pregnancy

Zang

Jiang

et al. 2022

IJMS

View full text Add to dashboard Cite

Litter size is an important indicator to measure the production capacity of commercial pigs. Spontaneous embryo loss is an essential factor in determining sow litter size. In early pregnancy, spontaneous embryo loss in porcine is as high as 20–30% during embryo implantation. However, the specific molecular mechanism underlying spontaneous embryo loss at the end of embryo implantation remains unknown. Therefore, we comprehensively used small RNA sequencing technology, bioinformatics analysis, and molecular experiments to determine the microRNA (miRNA) expression profile in the healthy and arresting embryo implantation site of porcine endometrium on day of gestation (DG) 28. A total of 464 miRNAs were identified in arresting endometrium (AE) and healthy endometrium (HE), and 139 differentially expressed miRNAs (DEMs) were screened. We combined the mRNA sequencing dataset from the SRA database to predict the target genes of these miRNAs. A quantitative real-time PCR assay identified the expression levels of miRNAs and mRNAs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed on differentially expressed target genes of DEMs, mainly enriched in epithelial development and amino acids metabolism-related pathways. We performed fluorescence in situ hybridization (FISH) and the dual-luciferase report gene assay to confirm miRNA and predicted target gene binding. miR-205 may inhibit its expression by combining 3′-untranslated regions (3′ UTR) of tubulointerstitial nephritis antigen-like 1 (TINAGL1). The resulting inhibition of angiogenesis in the maternal endometrium ultimately leads to the formation of arresting embryos during the implantation period. This study provides a reference for the effect of miRNA on the successful implantation of pig embryos in early gestation.

show abstract

“…The AceQ Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) and the miRNA Universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) were used to detect Quantitative real-time PCR (q-PCR) of mRNA and miRNA, respectively. The relative level of RNA expression was normalized to GAPDH and miR-17 [ 27 , 28 , 29 ] expression levels using the 2 −ΔΔCt method. Every sample was detected in triplicate.…”

Section: Methodsmentioning

confidence: 99%

SESN3 Inhibited SMAD3 to Relieve Its Suppression for MiR-124, Thus Regulating Pre-Adipocyte Adipogenesis

Lin

Zhao

Yan

et al. 2021

Genes

View full text Add to dashboard Cite

Sestrin-3, together with the other two members Sestrin-1 and Sestrin-2, belongs to the Sestrin family. The Sestrin protein family has been demonstrated to be involved in antioxidative, metabolic homeostasis, and even the development of nonalcoholic steatohepatitis (NASH). However, the adipogenic regulatory role of SESN3 in adipogenesis still needs to be further explored. In this study, we demonstrated SESN3 inhibited porcine pre-adipocyte proliferation, thus suppressing its adipogenesis. Meanwhile, SESN3 has been demonstrated to inhibit Smad3 thus protecting against NASH. Further, for our previous study, we found mmu-miR-124 involved in 3T3-L1 cell adipogenesis regulation. In this study, we also identified that ssc-miR-124 inhibited porcine pre-adipocyte proliferation, thus suppressing its adipogenesis, and the SMAD3 was an inhibitor of ssc-miR-124 by binding to its promoter. Furthermore, the ssc-miR-124 targeted porcine C/EBPα and GR and thus inhibited pre-adipocyte adipogenesis. In conclusion, SESN3 inhibited SMAD3, thus improving ssc-miR124, and then suppressed C/EBPα and GR to regulate pre-adipocytes adipogenesis.

show abstract

Identification of endogenous microRNA references in porcine serum for quantitative real-time PCR normalization

Cited by 6 publications

References 33 publications

Mean Expression Value Normalized and Absolute Quantified miR-21 Found to be a Potential Diagnostic and Prognostic Marker for IgA Nephropathy

Mean Expression Value Normalized and Absolute Quantified miR-21 Found to be a Potential Diagnostic and Prognostic Marker for IgA Nephropathy

Differential MicroRNA Expression in Porcine Endometrium Related to Spontaneous Embryo Loss during Early Pregnancy

SESN3 Inhibited SMAD3 to Relieve Its Suppression for MiR-124, Thus Regulating Pre-Adipocyte Adipogenesis

Contact Info

Product

Resources

About