Identification, amplification and characterization of miR-17-92 from canine tissue
ABSTRACT Recently, a novel group of genes encoding small RNA molecules, termed microRNAs (miRNAs), has been discovered to play a vital role in eukaryotic gene expression. Known to act in a post-transcriptional fashion, miRNAs can inhibit translation by binding to messenger RNA (mRNA) or by targeting mRNA for degradation. A search of genetic databases revealed significant conservation of miRNA genes between the domestic dog and the human. This finding suggests that expression patterns may also be conserved. Proof of principle experiments, including serial dilutions and sequencing, were performed to verify that primers made to amplify human mature miRNAs can be used to amplify canine miRNAs, providing that the mature sequences are conserved. TaqMan Real-time PCR techniques were used to isolate the first miRNA mature products from canine tissues. The expression levels of miR-17-3p, miR-17-5p, miR-18, miR-19a, miR-19b, miR-20, and miR-92 were evaluated in five canine tissues (heart, lung, brain, kidney, and liver) using the delta-delta Ct (critical threshold) method.
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- "Human and canine data were normalized compared with the endogenous reference RNU44, following Applied Biosystems indications (TaqMan miRNA assay n°4373384 – PE Applied Biosystems). We also tested RNU6B, an endogenous reference previously used in dogs (Boggs et al., 2007, 2008) but RNU44 was chosen as it proved to be more constant and reliable. Each was analysed in duplicate. "
ABSTRACT: Osteosarcoma (OS) is the most common primary malignant bone tumour in dogs and humans. MicroRNAs are short non-coding RNA molecules involved in post-transcriptional gene expression. Here, we compared the effects of miR-196a deregulation in human and canine OS cells after having observed a more uniform distribution and stronger down-expression in the human specimens. Cell response to miR-196a transfection was different in human and canine OS. A decreased proliferation rate was seen in human MG63 and 143B OS cells, while no appreciable changes occurred in canine DAN cells. Transient decrease of motility was highly remarkable and longer in MG63, concomitant with decreased levels of annexin1, a target of miR-196a promoting cell migration and invasion.Research in Veterinary Science 12/2014; 99. DOI:10.1016/j.rvsc.2014.12.017 · 1.51 Impact Factor
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ABSTRACT: A hereditary evaluation of Legg-Calve-Perthes Disease (LCPD) in the Miniature Pinscher (Min Pin) is proposed to assess the genetic contribution to disease presentation. Initial analyses include complex segregation analyses (CSA) to determine the mode of transmission of LCPD in the Min Pin, as well as heritability analyses to estimate the contribution of genetic and non-genetic (environmental) factors in the total population's phenotypic variance. Future studies are dependent on the results of the CSA: simple patterns of inheritance (i.e., autosomal recessive, autosomal dominant) require fewer dogs in association studies than complex traits (i.e., polygenic/multifactoral). Linkage studies (linkage analysis; linkage disequilibrium) draw on basic biological principles in that markers, or genetic "signposts" (e.g., microsatellites, single nucleotide polymorphisms (SNPs)) located near a disease-controlling gene will be co-inherited, or linked, with the disease phenotype. To determine linkage, there are two approaches we can take: classical linkage analysis (LA) which traces alleles through generations, and linkage disequilibrium (LD), which utilizes unrelated dogs that show the same phenotype.
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ABSTRACT: MicroRNAs (miRNAs) are 18-22-nt noncoding RNAs that are involved in post-transcriptional regulation of genes. Oncomirs, a subclass of miRNAs, include genes whose expression, or lack thereof, are associated with cancers. Until the last decade, the domestic dog was an underused model for the study of various human diseases that have genetic components. The dog exhibits marked genetic and physiologic similarity to the human, thereby making it an excellent model for study and treatment of various hereditary diseases. Furthermore, because the dog presents with distinct, spontaneously occurring mammary tumors, it may serve as a model for genetic analysis and treatments of humans with malignant breast tumors. Because miRNAs have been found to act as both tumor suppressors and oncogenes in several different cancers, expression patterns of ten miRNAs (miR-15a, miR-16, miR-17-5p, miR-21, miR-29b, miR-125b, miR-145, miR-155, miR-181b, let-7f) known to be associated with human breast cancers were compared to malignant canine mammary tumors (n = 6) and normal canine mammary tissue (n = 10). Resulting data revealed miR-29b and miR-21 to have a statistically significant (p < 0.05 by MANOVA analysis) upregulation in cancerous samples. The ten canine miRNAs follow the same pattern of expression as in the human, except for miR-145 which does not show a difference in expression between the normal and cancerous canine samples. In addition, when analyzed according to specific cancer phenotypes, miR-15a and miR-16 show a significant downregulation in canine ductal carcinomas while miRsR-181b, -21, -29b, and let-7f show a significant upregulation in canine tubular papillary carcinomas.Mammalian Genome 08/2008; 19(7-8):561-9. DOI:10.1007/s00335-008-9128-7 · 2.88 Impact Factor