Project description:To investigate the molecular mechanism of the hypopigmentation observed in Dicer KO mice, Dicer knockdown was realised in vitro in normal C57BL/6 mouse melanocyte Melan-a cells. Transient transfection of Melan-a cells with siRNA directed against Dicer reduced Dicer protein levels to approximately 40% of that of siScr Melan-a cells 24 hours after transfection. We analyzed the miRnome of Melan-a cells 24 and 48 hours after transfection with siDicer or siScr to understand the molecular mechanisms involved in Dicer-dependent melanocyte migration.
Project description:To investigate the molecular mechanism of the hypopigmentation observed in Dicer KO mice, Dicer knockdown was realised in vitro in normal C57BL/6 mouse melanocyte Melan-a cells. Transient transfection of Melan-a cells with siRNA directed against Dicer reduced Dicer protein levels to approximately 40% of that of siScr Melan-a cells 48 hours after transfection. We analyzed the transcriptome of Melan-a cells 48 hours after transfection with siDicer or siScr to understand the molecular mechanisms involved in Dicer-dependent melanocyte migration.
Project description:To investigate the molecular mechanism of the hypopigmentation observed in Dicer KO mice, Dicer knockdown was realised in vitro in normal C57BL/6 mouse melanocyte Melan-a cells. Transient transfection of Melan-a cells with siRNA directed against Dicer reduced Dicer protein levels to approximately 40% of that of siScr Melan-a cells 24 hours after transfection. We analyzed the transcriptome of Melan-a cells 24 hours after transfection with siDicer or siScr to understand the molecular mechanisms involved in Dicer-dependent melanocyte migration.
Project description:DICER has a well-characterized role in the processing of microRNAs (miRNAs) and small interfering RNAs (siRNA) that are important for post-transcriptional gene regulation. Emerging evidence suggests that DICER also has several non-canonical functions beyond miRNA/siRNA biogenesis, for example in transcriptional gene silencing at the chromatin level, as well as in RNA degradation and maintenance of genomic integrity. We have shown that the function of DICER in germ cells is essential for normal spermatogenesis; male mice lacking DICER in postnatal male germ cells are infertile due to severe defects in haploid differentiation. To better understand the function of DICER in male germ cells, we immunoprecipitated DICER from juvenile mouse testes and performed mass spectrometric analysis to identify DICER-interacting proteins.
Project description:MicroRNA regulates gene expression. In this experiment, we determined the gene expression changes by the microRNA expression status using Dicer protein nulll. Wildtype, and intermediate expression. Dicer gene disrupted HCT116 colon cancer cells were transduced with tetracycline regulated Dicer protein expression. To determine the gene expression changes dependent on the Dicer protein levels, we used Dicer disrupted cells, Dicer wild-type cells, and cells with intermediate expression level of Dicer protein for the cDNA microarrya analyses.
Project description:Analysis of mRNA changes in HeLa cells following Ago2 or Dicer depletion. Dicer, a cytoplasmic RNase III, generates the mature form of small RNAs including microRNA. Ago2 is a component of an effector complex of microRNA. Keywords: gene expression array-based (RNA / in situ oligonucleotide)