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PLoS Genet. 2016 Jun 2;12(6):e1006095. doi: 10.1371/journal.pgen.1006095. eCollection 2016 Jun.

Endogenous Mouse Dicer Is an Exclusively Cytoplasmic Protein.

Author information

1
Mouse Biology Unit, European Molecular Biology Laboratory (EMBL), Monterotondo, Italy.
2
MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
3
Wellcome Trust Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, Scotland, United Kingdom.
4
Genomics Core Facility, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.

Abstract

Dicer is a large multi-domain protein responsible for the ultimate step of microRNA and short-interfering RNA biogenesis. In human and mouse cell lines, Dicer has been shown to be important in the nuclear clearance of dsRNA as well as the establishment of chromatin modifications. Here we set out to unambiguously define the cellular localization of Dicer in mice to understand if this is a conserved feature of mammalian Dicer in vivo. To this end, we utilized an endogenously epitope tagged Dicer knock-in mouse allele. From primary mouse cell lines and adult tissues, we determined with certainty by biochemical fractionation and confocal immunofluorescence microscopy that endogenous Dicer is exclusively cytoplasmic. We ruled out the possibility that a fraction of Dicer shuttles to and from the nucleus as well as that FGF or DNA damage signaling induce Dicer nuclear translocation. We also explored Dicer localization during the dynamic and developmental context of embryogenesis, where Dicer is ubiquitously expressed and strictly cytoplasmic in all three germ layers as well as extraembryonic tissues. Our data exclude a direct role for Dicer in the nuclear RNA processing in the mouse.

PMID:
27254021
PMCID:
PMC4890738
DOI:
10.1371/journal.pgen.1006095
[Indexed for MEDLINE]
Free PMC Article

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