Creation of a Large Genomic Deletion at the T-Cell Antigen Receptor β-Subunit Locus in Mouse Embryonic Stem Cells by Gene Targeting

Recently it has become possible to introduce predesigned mutations into a given gene in the mouse germ line by homologous recombination in embryonic stem cells. The mutations are usually introduced by inserting the neomycin phosphotransferase gene into an exon of a particular gene. Here we describe...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1991-04, Vol.88 (8), p.3084-3087
Main Authors: Mombaerts, Peter, Clarke, Alan R., Hooper, Martin L., Tonegawa, Susumu
Format: Article
Language:English
Subjects:
Animals
Biological and medical sciences
Blotting, Southern
Cell Line
Cell lines
Chromosomes
DNA Mutational Analysis
DNA probes
Fundamental and applied biological sciences. Psychology
Genes
Genetic loci
Genomics
Homologous recombination
Mice
Molecular and cellular biology
Molecular genetics
Mutagenesis. Repair
Plasmids
Receptors, Antigen, T-Cell - genetics
Receptors, Antigen, T-Cell, alpha-beta
Recombination, Genetic
Restriction Mapping
Stem cells
Stem Cells - physiology
T cell antigen receptors
Transfection
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Summary:Recently it has become possible to introduce predesigned mutations into a given gene in the mouse germ line by homologous recombination in embryonic stem cells. The mutations are usually introduced by inserting the neomycin phosphotransferase gene into an exon of a particular gene. Here we describe an extension of this method that can result in at least a 15-kilobase-long deletion. The deletion created in the present work encompasses one of the two diversity gene segments of the mouse T-cell receptor β-subunit locus, 10 out of the 12 joining gene segments, and both constant gene segments. This strategy is a valuable alternative to sequential targeting of multiple genes forming a gene cluster, could simplify the construction of plasmids to be used for targeting, and could be the solution for inactivating small genes that have eluded conventional targeting approaches.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.88.8.3084