Dicer-deficient mouse embryonic stem cells are defective in differentiation and centromeric silencing.

By C. Kanellopoulou, Stefan A. Muljo, A. Kung, Shridar Ganesan, R. Drapkin, T. Jenuwein, D. Livingston, K. Rajewsky. Genes & Development, 2005

1,357 citations recorded by the source corpus.

Dicer is the enzyme that cleaves double-stranded RNA (dsRNA) into 21-25-nt-long species responsible for sequence-specific RNA-induced gene silencing at the transcriptional, post-transcriptional, or translational level. We disrupted the dicer-1 (dcr-1) gene in mouse embryonic stem (ES) cells by conditional gene targeting and generated Dicer-null ES cells. These cells were viable, despite being completely defective in RNA interference (RNAi) and the generation of microRNAs (miRNAs). However, the mutant ES cells displayed severe defects in differentiation both in vitro and in vivo. Epigenetic silencing of centromeric repeat sequences and the expression of homologous small dsRNAs were markedly reduced. Re-expression of Dicer in the knockout cells rescued these phenotypes. Our data suggest that Dicer participates in multiple, fundamental biological processes in a mammalian organism, ranging from stem cell differentiation to the maintenance of centromeric heterochromatin structure and centromeric silencing.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/fbf9c41ad859fe641b6b3d5f0e0fe09f692cec0d

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Citations in the corpus

• How is pluripotency determined and maintained? (2007, background): “5 Defective Retarded/ compensated Normal Defective Restored Bernstein et al., 2003; Kanellopoulou et al., 2005; Murchison et al., 2005 Chromatin remodeling/Histone exchange Snf2b (Brg1, Smarca4) Die at E4.”
• Origins and Mechanisms of miRNAs and siRNAs (2009, background): “Surprisingly, mouse cells carrying a null allele of Dicer can still assemble siRNAs into functional RISCs (Kanellopoulou et al., 2005; Murch-ison et al., 2005), indicating that Dicer is not required for RISC loading in mammals.”
• Chromatin in pluripotent embryonic stem cells and differentiation (2006, background): “The relevance of this mechanism is indicated by the observation that differentiation is perturbed in ES cells that lack Dicer, the enzyme that is responsible for the cleavage of double-stranded RNAs into small interfering (si)RNA”
• RNA meets chromatin. (2005, neutral): “cells (Kanellopoulou et al. 2005).”
• Transposable elements and the epigenetic regulation of the genome (2007, background): “These mutants also have centromeric condensation and differentiation defect”
• Epigenetic inheritance during the cell cycle (2009, background): “Whereas experimental evidence substantiates this model in yeast, whether RNAi is involved in heterochromatin maintenance in mammals is unclea”
• DNA damage-induced alterations in chromatin contribute to genomic integrity and age-related changes in gene expression (2008, positive): “…and the pan-class III HDAC (sirtuin) inhibitors NAM and sirtinol caused desilencing of major satellite repeats (Figure 1F and Figures S1E and S1F) (Kanellopoulou et al., 2005), whereas no significant increase in repeat transcripts was seen after stable knockdown of SIRT1 (Figures S1C and S1D and…”
• MicroRNA and cancer (2012, background): “For example, the let-7 target HMGA2 is a repressor of the key inducers of senescence p16 and p19, products of the INK4A locus often mutated in cancers (Lee and Dutta, 2007; Nishino et al., 2008; Sherr, 1996). p16 is a specific inhibitor of CDK4/6 and is also repressed by miR24, which is down-regulat”
• Control of Embryonic Stem Cell State (2011, background): “…in this family have been implicated in cell proliferation (He et al., 2005; O’Donnell et al., 2005; Wang et al., 2008) and have been shown to rescue the proliferation defects observed in miRNA-deficient ES cells (Kanellopoulou et al., 2005; Murchison et al., 2005; Wang et al., 2007, 2008).”
• Mouse ES cells express endogenous shRNAs, siRNAs, and other Microprocessor-independent, Dicer-dependent small RNAs. (2008, methodology): “Repeat element and NCBI refSeq gene annotations were obtained from UCSC (Jurka 2000; Karolchik et al. 2003; Pruitt et al. 2005).”
• MicroRNAs: key regulators of stem cells (2009, background): “This shows that miRNAs and/or endo-siRNAs might function in ES cell differentiatio”
• Identification and characterization of two novel classes of small RNAs in the mouse germline: retrotransposon-derived siRNAs in oocytes and germline small RNAs in testes. (2006, background): “In support of this view, an increase of retrotransposon transcripts has been reported in Dicer-deficient embryonic stem (ES) cells and Dicerknockdown preimplantation embryos in mouse (Svoboda et al. 2004a; Kanellopoulou et al. 2005).”
• Linking DNA methylation and histone modification: patterns and paradigms (2009, background): “The resulting RNA-induced silencing complex (RISC) is then specifically targeted back to pericentromeric regions where it probably recruits SUV39H1 and SUV39H2, which are the most important components in this heterochromatin pathwa”