Document Detail


Efficient modification of human chromosomal alleles using recombination-proficient chicken/human microcell hybrids.
MedLine Citation:
PMID:  8563756     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Targeted modification of human chromosomal alleles by homologous recombination is a powerful approach to study gene function, but gene targeting in mammalian cells is an inefficient process. In contrast, gene targeting in a chicken pre-B cell line, DT40, is highly efficient. We have transferred human chromosome 11 into DT40 cells by microcell fusion, and find that the resulting hybrids are recombination-proficient. In these cells, targeting efficiencies into the chicken ovalbumin locus were > 90% and into the human beta-globin and Ha-ras loci were 10-15%. These modified human chromosomes can be transferred subsequently to mammalian cells for functional tests. This chromosome shuttle system allows for the efficient homologous modification of human chromosomal genes, and for subsequent phenotypic analyses of the modified alleles in different mammalian cell types.
Authors:
E S Dieken; E M Epner; S Fiering; R E Fournier; M Groudine
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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:  Nature genetics     Volume:  12     ISSN:  1061-4036     ISO Abbreviation:  Nat. Genet.     Publication Date:  1996 Feb 
Date Detail:
Created Date:  1996-03-05     Completed Date:  1996-03-05     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  9216904     Medline TA:  Nat Genet     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  174-82     Citation Subset:  IM    
Affiliation:
Division of Molecular Medicine, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104, USA.
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MeSH Terms
Descriptor/Qualifier:
Alleles*
Animals
B-Lymphocytes
Base Sequence
Cell Fusion
Cell Line
Chickens
Chromosomes, Human, Pair 11
Gene Targeting / methods*
Genes, ras / genetics
Globins / genetics
Humans
Hybrid Cells*
Leukemia, Erythroblastic, Acute
Mice
Molecular Sequence Data
Ovalbumin / genetics
RNA, Messenger / analysis
Recombination, Genetic / genetics*
Tumor Cells, Cultured
Grant Support
ID/Acronym/Agency:
CA54337/CA/NCI NIH HHS; DK44746/DK/NIDDK NIH HHS; GM26449/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/RNA, Messenger; 9004-22-2/Globins; 9006-59-1/Ovalbumin

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


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