title: Molecular quantification of canine specific microRNA species.
abstract: 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.
authors.affiliation: Institute of Veterinary Pathology, Freie Universität Berlin, Robert-von-Ostertagstraße 15, 14163 Berlin, Germany.
authors.forename: M-C
authors.initials: MC
authors.lastname: von Deetzen
authors.forename: B
authors.initials: B
authors.lastname: Schmeck
authors.forename: A D
authors.initials: AD
authors.lastname: Gruber
authors.forename: R
authors.initials: R
authors.lastname: Klopfleisch
mesh_terms: D000818:Animals; D004285:Dogs / Q000235:genetics* / Q000378:metabolism*; D005456:Fluorescent Dyes; D005786:Gene Expression Regulation / Q000502:physiology*; D005819:Genetic Markers; D035683:MicroRNAs / Q000235:genetics*; D020537:RNA, Small Nucleolar / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
chemical_list: D005456:Fluorescent Dyes; D005819:Genetic Markers; D035683:MicroRNAs; D020537:RNA, Small Nucleolar
keywords: Canine; Gene expression; Housekeeping gene; MicroRNA; Quantitative PCR; Small nucleolar RNA