Document Detail


Identification and targeting of the ROSA26 locus in human embryonic stem cells.
MedLine Citation:
PMID:  18037879     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The derivation of human embryonic stem (hES) cells has opened new avenues for studies on human development and provided a potential source of cells for replacement therapy. To reveal the full potential of hES cells, it would be advantageous to be able to genetically alter them as is routinely done with mouse ES cells through homologous recombination. The mouse Rosa26 locus is particularly useful for genetic modification as it can be targeted with high efficiency and is expressed in most cell types tested. Here we report the identification of the human homolog of the mouse Rosa26 locus. We demonstrate targeting of a red-fluorescent protein (tdRFP) cDNA to this locus through homologous recombination and expression of this targeted reporter in multiple hES cell-derived lineages. Through recombinase-mediated cassette exchange, we show replacement of the tdRFP cDNA with other cDNAs, providing a cell line in which transgenes can be readily introduced into a broadly expressed locus.
Authors:
Stefan Irion; Hervé Luche; Paul Gadue; Hans Joerg Fehling; Marion Kennedy; Gordon Keller
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2007-11-25
Journal Detail:
Title:  Nature biotechnology     Volume:  25     ISSN:  1546-1696     ISO Abbreviation:  Nat. Biotechnol.     Publication Date:  2007 Dec 
Date Detail:
Created Date:  2007-12-10     Completed Date:  2008-02-22     Revised Date:  2010-09-29    
Medline Journal Info:
Nlm Unique ID:  9604648     Medline TA:  Nat Biotechnol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1477-82     Citation Subset:  IM    
Affiliation:
University Health Network, McEwen Centre for Regenerative Medicine, 101 College Street, Toronto, ON, M5G 1L7, Canada.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cells, Cultured
Embryonic Stem Cells / cytology,  physiology*
Gene Targeting / methods*
Humans
Mice
Proteins / genetics*,  metabolism*
Recombinant Proteins / metabolism*
Transfection / methods*
Grant Support
ID/Acronym/Agency:
HL8062/HL/NHLBI NIH HHS; P20GM075019/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Gt(ROSA)26Sor protein, mouse; 0/Proteins; 0/Recombinant Proteins

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


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