Molecular quantification of canine specific microRNA species

MicroRNAs (miRNAs) belong to the heterogeneous group of non-coding, regulatory RNA molecules. They play an important role in physiological functions, especially during tissue development. Moreover, their cell-type specific expression levels are also thought to be altered in association with differen...

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Bibliographic Details
Published in:Research in veterinary science 2013-10, Vol.95 (2), p.562-568
Main Authors: von Deetzen, M.-C., Schmeck, B., Gruber, A.D., Klopfleisch, R.
Format: Article
Language:English
Subjects:
Animals
Biomedical research
Canine
complementary DNA
Dogs
Dogs - genetics
Dogs - metabolism
essential genes
Fluorescent Dyes
Gene expression
Gene Expression Regulation - physiology
Genetic Markers
Housekeeping gene
humans
metabolic diseases
Methods
mice
Microbiology
MicroRNA
MicroRNAs - genetics
Quantitative PCR
quantitative polymerase chain reaction
Real-Time Polymerase Chain Reaction - veterinary
Reverse Transcriptase Polymerase Chain Reaction - veterinary
RNA, Small Nucleolar - genetics
Small nucleolar RNA
Studies
Veterinary medicine
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Summary:MicroRNAs (miRNAs) belong to the heterogeneous group of non-coding, regulatory RNA molecules. They play an important role in physiological functions, especially during tissue development. Moreover, their cell-type specific expression levels are also thought to be altered in association with different pathological conditions, including metabolic disorders and cancer. However, the vast majority of molecular assays for their quantification have been established for humans and mice, with no reports on specific research tools for dogs. Here canine-specific SYBRGreen based quantitative, real-time PCR assays are introduced for the quantification of 21 miRNAs and five small nucleolar RNA (snoRNA). The assay includes a time-saving and economical universal one-tube cDNA synthesis approach which allows for a more robust relative quantification than the commonly used multiple specific reverse transcriptions. GeNorm and NormFinder analysis were used for the evaluation of potential housekeeping genes and identified miRNAs to be more stable housekeeping genes by trend.
ISSN:0034-5288
1532-2661
DOI:10.1016/j.rvsc.2013.06.012