Creation of a large genomic deletion at the T-cell antigen receptor beta-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 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 beta-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.
Mombaerts
P
P
Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge 02139.
Clarke
A R
AR
Hooper
M L
ML
Tonegawa
S
S
D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D004252:DNA Mutational Analysis; D051379:Mice; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011995:Recombination, Genetic; D015183:Restriction Mapping; D013234:Stem Cells / Q000502:physiology; D014162:Transfection
D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta