Characterization of Dicer-deficient murine embryonic stem cells

Murchison, E. P., Partridge, J. F., Tam, O. H., Cheloufi, S., Hannon, G. J. (August 2005) Characterization of Dicer-deficient murine embryonic stem cells. Proceedings of the National Academy of Sciences of the United States of America, 102 (34). pp. 12135-40. ISSN 0027-8424

[img]
Preview
PDF (Paper)
Hannon PNAS 2005b.pdf - Published Version

Download (569Kb) | Preview
URL: http://www.ncbi.nlm.nih.gov/pubmed/16099834
DOI: 10.1073/pnas.0505479102

Abstract

Dicer is an RNase III-family nuclease that initiates RNA interference (RNAi) and related phenomena by generation of the small RNAs that determine the specificity of these gene silencing pathways. We have previously shown that Dicer is essential for mammalian development, with Dicer-deficient mice dying at embryonic day 7.5 with a lack of detectable multipotent stem cells. To permit a more detailed investigation of the biological roles of Dicer, we have generated embryonic stem cell lines in which their single Dicer gene can be conditionally inactivated. As expected, Dicer loss compromises maturation of microRNAs and leads to a defect in gene silencing triggered by long dsRNAs. However, the absence of Dicer does not affect the ability of small interfering RNAs to repress gene expression. Of interest, Dicer loss does compromise the proliferation of ES cells, possibly rationalizing the phenotype previously observed in Dicer-null animals. Dicer loss also affects the abundance of transcripts from mammalian centromeres but does so without a pronounced affect on histone modification status at pericentric repeats or methylation of centromeric DNA. These studies provide a conditional model of RNAi deficiency in mammals that will permit the dissection of the biological roles of the RNAi machinery in cultured mammalian cells.

Item Type: Paper
Uncontrolled Keywords: Animals Blotting Northern Blotting Southern Blotting Western Cell Proliferation Chromatin Immunoprecipitation DNA Primers DNA Complementary genetics Flow Cytometry Gene Expression Regulation Mice MicroRNAs metabolism RNA Interference Ribonuclease III deficiency Stem Cells cytology Transfection
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > dicer
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > dicer

organism description > animal > developmental stage > embryo
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene silencing
organism description > animal > mammal > rodent > mouse
therapies > stem cells
CSHL Authors:
Communities: CSHL labs > Hannon lab
Watson School > Publications
Depositing User: Editor Margaret Fantz
Date: 23 August 2005
Date Deposited: 10 Apr 2012 19:07
Last Modified: 11 Jan 2018 17:55
PMCID: PMC1185572
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26130

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving