Document Detail


Reversible immortalization of mammalian cells mediated by retroviral transfer and site-specific recombination.
MedLine Citation:
PMID:  8799138     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A procedure of reversible immortalization of primary cells was devised by retrovirus-mediated transfer of an oncogene that could be subsequently excised by site-specific recombination. This study focused on the early stages of immortalization: global induction of proliferation and life span extension of cell populations. Comparative analysis of Cre/LoxP and FLP/FRT recombination in this system indicated that only Cre/LoxP operates efficiently in primary cells. Pure populations of cells in which the oncogene is permanently excised were obtained, following differential selection of the cells. Cells reverted to their preimmortalized state, as indicated by changes in growth characteristics and p53 levels, and their fate conformed to the telomere hypothesis of replicative cell senescence. By permitting temporary and controlled expansion of primary cell populations without retaining the transferred oncogene, this strategy may facilitate gene therapy manipulations of cells unresponsive to exogenous growth factors and make practical gene targeting by homologous recombination in somatic cells. The combination of retroviral transfer and site-specific recombination should also extend gene expression studies to situations previously inaccessible to experimentation.
Authors:
K A Westerman; P Leboulch
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  93     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1996 Aug 
Date Detail:
Created Date:  1996-10-31     Completed Date:  1996-10-31     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  8971-6     Citation Subset:  IM    
Affiliation:
Harvard-Massachusetts Institute of Technology Division of Health Sciences and Technology, Cambridge 02139, USA.
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MeSH Terms
Descriptor/Qualifier:
3T3 Cells
Animals
Cattle
Cell Aging
Cell Line, Transformed
Cell Transformation, Viral*
DNA Nucleotidyltransferases
Gene Transfer Techniques*
Genetic Vectors
Humans
Integrases
Kidney / cytology
Mice
Muscle, Smooth, Vascular / cytology
Rabbits
Recombination, Genetic*
Simian virus 40 / genetics*
Skin / cytology
Telomere / metabolism
Transduction, Genetic
Viral Proteins*
Grant Support
ID/Acronym/Agency:
HL48374-01/HL/NHLBI NIH HHS; HL55435-01/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Viral Proteins; EC 2.7.7.-/Cre recombinase; EC 2.7.7.-/DNA Nucleotidyltransferases; EC 2.7.7.-/FLP recombinase; EC 2.7.7.-/Integrases
Comments/Corrections

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


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