Document Detail


Visualizing loss of heterozygosity in living mouse cells and tissues.
MedLine Citation:
PMID:  18708075     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Loss of heterozygosity (LOH) in somatic cells can contribute to the genesis of cancer, but little is known about the frequency with which LOH occurs in normal cells of the body. To detect LOH in situ, we studied mouse shYFP embryonic stem (ES) cells and cells of the intestinal epithelia derived from these ES cells. shYFP ES cells are heterozygous at the ROSA26 locus. One copy of the locus carries a gene encoding a yellow fluorescent protein (YFP), while the other copy harbors an shRNA gene that produces a short hairpin RNA (shRNA) molecule that causes degradation of YFP mRNA. Nearly all cells in shYFP populations were faintly fluorescent, but brightly fluorescent cells arose at a rate of approximately 10(-5)bright cells/generation. Bright cells lacked the gene encoding the shRNA and contained two copies of the YFP gene. Comparison of these results to previous data on LOH in ES cells that lacked interfering shRNA showed that LOH in shYFP cells was not influenced by the presence of the shRNA. Bright cells were also seen in intestinal villi of chimeric mice made by injecting blastocysts with shYFP cells. These data demonstrate that this approach can detect LOH and suggest that it will allow detection of LOH in a broad array of tissues and cell types in transgenic mice made from shYFP cells.
Authors:
Jared M Fischer; James R Stringer
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2008-07-30
Journal Detail:
Title:  Mutation research     Volume:  645     ISSN:  0027-5107     ISO Abbreviation:  Mutat. Res.     Publication Date:  2008 Oct 
Date Detail:
Created Date:  2008-10-20     Completed Date:  2009-01-06     Revised Date:  2011-04-28    
Medline Journal Info:
Nlm Unique ID:  0400763     Medline TA:  Mutat Res     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1-8     Citation Subset:  IM    
Affiliation:
University of Cincinnati, Department of Molecular Genetics, Biochemistry and Microbiology, Cincinnati, OH 45267-0524, USA. fischerjm@gmail.com
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MeSH Terms
Descriptor/Qualifier:
Animals
Base Sequence
DNA Primers / genetics
Embryonic Stem Cells / metabolism
Epithelial Cells / metabolism
Intestine, Small / cytology,  metabolism
Loss of Heterozygosity*
Luminescent Proteins / genetics
Mice
Mice, Transgenic
Microscopy, Fluorescence
RNA Interference
RNA, Messenger / genetics,  metabolism
Grant Support
ID/Acronym/Agency:
ES07250-19/ES/NIEHS NIH HHS
Chemical
Reg. No./Substance:
0/DNA Primers; 0/Luminescent Proteins; 0/RNA, Messenger

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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