Project description:A majority of genetically modified mice carry passenger mutations, originating from the 129-derived embryonic stem (ES) cells, near the targeted gene. Unintended retention of these mutations can introduce confounding phenotypes and affect conclusions. We show that Ackr1-/-129ES mice retained approximately 6 Mb of 129 ES cells-derived genomic material in chromosome 1 near Ackr1 in bone marrow derived monocytes (Mono), nucleated erythroblasts (NECs), and polymorphonucleated neutrophils (PMN), notably affecting the expression of PYHIN genes.
Project description:Experiment 2 - MiRNA mimics have a length and passenger strand specific effect Wildtype and mutant variants of miR-155 and negative control miRNA mimics were transfected to address the phenotype
Project description:1 year-old male Tenascin-C deficient 129/SV mice with the targeted insertion of a beta-lactamase cassette in the NcoI site of exon 2 of the TNC gene (Forsberg, E., Hirsch, E., Frohlich, L., Meyer, M., Ekblom, P., Aszodi, A., Werner, S. & Fassler, R. (1996) Proc. Natl. Acad. Sci U. S. A 93, 6594-6599) were used for the study. Animals were derived from the original strain and back-crossed with wildtype 129/SV mice (Institut für Labortierkunde, University of Zurich). Animals were acclimatized to housing in single cages for one week. Soleus muscle was harvested, total RNA isolated and subjected to transcript profiling with custom microarrays with custom-designed ATLASTM cDNA nylon filter as described (Dapp, C., Schmutz, S., Hoppeler, H. & Fluck, M. (2004) Physiol Genomics 20, 97-107). Keywords: Transcriptome
Project description:1 year-old male Tenascin-C deficient 129/SV mice with the targeted insertion of a beta-lactamase cassette in the NcoI site of exon 2 of the TNC gene (Forsberg, E., Hirsch, E., Frohlich, L., Meyer, M., Ekblom, P., Aszodi, A., Werner, S. & Fassler, R. (1996) Proc. Natl. Acad. Sci U. S. A 93, 6594-6599) were used for the study. Animals were derived from the original strain and back-crossed with wildtype 129/SV mice (Institut für Labortierkunde, University of Zurich). Animals were acclimatized to housing in single cages for one week followed by 7 days of hindlimb suspension and one day of reloading. Soleus muscle was harvested, total RNA isolated and subjected to transcript profiling with custom microarrays with custom-designed ATLASTM cDNA nylon filter as described (Dapp, C., Schmutz, S., Hoppeler, H. & Fluck, M. (2004) Physiol Genomics 20, 97-107). Keywords: Transcriptome
Project description:Primary cilium serves as a cellular “antenna” to sense environmental signals. Ciliogenesis requires the removal of CP110 to convert the mother centriole into the basal body. Actin dynamics is also critical for cilia formation. How these distinct processes are properly regulated remains unknown. Here we show that miR-129-3p, a microRNA conserved in the vertebrates, controlled cilia assembly by down-regulating both CP110 and four proteins critical for actin dynamics, Arp2, Toca1, abLIM1, and abLIM3. Consistently, blocking miR-129-3p repressed cilia formation in cultured mammalian cells, whereas its overexpression potently induced ciliogenesis in proliferating cells and extraordinary cilia elongation. Moreover, inhibition of miR-129-3p in zebrafish embryos suppressed cilia assembly in the Kupffer’s vesicle and pronephric duct, leading to developmental abnormalities including curved body, pericardial oedema, and randomised left-right patterning. Our results thus unravel a novel mechanism that orchestrates both the centriole-to-basal body transition and subsequent cilia assembly via microRNA-mediated posttranscriptional regulations. We want to find the targets of miR-129-3p by overexpressing miR-129-3p oligo or control oligo in hTERT-RPE1 cells. Through microarray analysis we could check the downregulated genes and these genes might be the targets of miR-129-3p.
Project description:Experiment 2 - MiRNA mimics have a length and passenger strand specific effect Wildtype and mutant variants of miR-155 and negative control miRNA mimics were transfected to address the phenotype Cells were transfected with mimics and samples were collected in duplicate (except 23mer miR-155 wild type mimic and 23mer negative control mimic)