Identification of Reference Genes for Relative Quantification of Circulating MicroRNAs in Bovine Serum

Title
Identification of Reference Genes for Relative Quantification of Circulating MicroRNAs in Bovine Serum
Author(s)
최인호배인선[배인선]정기용[정기용]김태일[김태일]이종민[이종민]최화식[최화식]조용무[조용무]김상훈[김상훈]
Keywords
QRT-PCR; RT-PCR; BIOMARKERS; EXPRESSION; PLASMA; NORMALIZATION; TISSUES; CANCER; MECHANISM; DISEASES
Issue Date
201503
Publisher
PUBLIC LIBRARY SCIENCE
Citation
PLOS ONE, v.10, no.3
Abstract
Circulating microRNAs in body fluids have been implicated as promising biomarkers for physiopathology disorders. Currently, the expression levels of circulating microRNAs are estimated by reverse transcription quantitative real-time polymerase chain reaction. Use of appropriate reference microRNAs for normalization is critical for accurate microRNA expression analysis. However, no study has systematically investigated reference genes for evaluating circulating microRNA expression in cattle. In this study, we describe the identification and characterization of appropriate reference microRNAs for use in the normalization of circulating microRNA levels in bovine serum. We evaluated the expression stability of ten candidate reference genes in bovine serum by using reverse transcription quantitative real-time polymerase chain reaction. Data were analyzed using geNorm, NormFinder, and Best-Keeper statistical algorithms. The results consistently showed that a combination of miR-93 and miR-127 provided the most stably expressed reference. The suitability of these microRNAs was validated, and even when compared among different genders or breeds, the combination of miR-93 and miR-127 was ranked as the most stable microRNA reference. Therefore, we conclude that this combination is the optimal endogenous reference for reverse transcription quantitative real-time polymerase chain reaction-based detection of microRNAs in bovine serum. The data presented in this study are crucial to successful biomarker discovery and validation for the diagnosis of physiopathological conditions in cattle.
URI
http://hdl.handle.net/YU.REPOSITORY/33042http://dx.doi.org/10.1371/journal.pone.0122554
ISSN
1932-6203
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생명공학부 > 생명공학부 > Articles
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