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

• 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
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.88.8.3084

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.