Project description:We investigated the effect of Dgcr8-homozygous mutation on microRNA expression profile in mouse embryonic stem cells. MicroRNA expression was substantially impaired, indicating a pivotal role of DGCR8 in microRNA biogenesis. MicroRNA expression profile was compared between Dgcr8-homozygous mutant ES cells and wild-type ES cells
Project description:We investigated the effect of Dgcr8-homozygous mutation on microRNA expression profile in mouse embryonic stem cells. MicroRNA expression was substantially impaired, indicating a pivotal role of DGCR8 in microRNA biogenesis.
Project description:Two independent BayGenomics mouse embryonic stem cell lines (XH157 and XG058), each bearing a gene trap between exons 9 and 10 of the Dgcr8 locus (ENSMUST00000115633, Ensembl v53) were further mutagenised by the targetted insertion of a second Dgcr8 gene trap cassette. Consequently two independent heterozygous and two independent homozygous mutant Dgcr8 cells were derived (Dgcr8tm1,gt1/+, Dgcr8tm1,gt2/+, Dgcr8gt1/tm1 and Dgcr8gt2/tm1). Next generation sequencing was used to profile the small RNAs expressed in these cells together with wild type (E14) cells to determine the effect of these mutations on miRNA expression.
Project description:These experiments were conducted as part of a study to derive the targets of miRNAs. Two independent BayGenomics mouse embryonic stem cell lines (XH157 and XG058), each bearing a gene trap between exons 9 and 10 of the Dgcr8 locus (ENSMUST00000115633, Ensembl v53) were further mutagenised by the targetted insertion of a second Dgcr8 gene trap cassette. Consequently two independent heterozygous and two independent homozygous mutant Dgcr8 cells were derived (Dgcr8tm1,gt1/+, Dgcr8tm1,gt2/+, Dgcr8gt1/tm1 and Dgcr8gt2/tm1). A comparison of the expression profiles of the heterozygous cells to the Dgcr8 depleted cells allows the investigation of the broad role of miRNAs in mouse ES cells. Subsequently individual miRNA mimics were reintroduced into the Dgcr8-depleted background. The cells were allowed to recover for 10, 20 or 44 hours. The effect of the miRNA on the mRNA expression of the cells was assessed through the comparison of the expression profile of the transfected cells to that of cells transfected with a control mimic, which had recovered over the same period. This allows the roles of individual miRNAs to be investigated.
Project description:These experiments were conducted as part of a study to derive the targets of miRNAs (See also E-MTAB-418). Two independent BayGenomics mouse embryonic stem cell lines (XH157 and XG058), each bearing a gene trap between exons 9 and 10 of the Dgcr8 locus (ENSMUST00000115633, Ensembl v53) were further mutagenised by the targetted insertion of a second Dgcr8 gene trap cassette. Consequently two independent heterozygous and two independent homozygous mutant Dgcr8 cells were derived (Dgcr8tm1,gt1/+, Dgcr8tm1,gt2/+, Dgcr8gt1/tm1 and Dgcr8gt2/tm1). Subsequently individual miRNA mimics were reintroduced into the Dgcr8-depleted background. In this set of experiments, cells were allowed to recover for 10 hours. The effect of the miRNA on the mRNA expression of the cells was assessed through the comparison of the expression profile of the transfected cells to that of cells transfected with a control mimic, which had recovered over the same period. This allows the roles of individual miRNAs to be investigated.
Project description:We performed Fluidigm C1 single cell sequencing analysis of wild-type and microRNA deficient (Dgcr8 knockout) mouse embryonic stem cells mock treated or transfected with either miR-294 or let-7.
Project description:To describe the protein profile in hippocampus, colon and ileum tissue’ changing after the old faeces transplants, we adopted a quantitative label free proteomics approach.
