Targeted removal of the mu switch region from mouse hybridoma cells. A test of its role in gene expression in the endogenous IgH locus.
The switch regions adjoining the DNA encoding the Ig heavy chain constant regions have been implicated in gene expression as well as isotype switching, in that transgenic mice express switch-containing transgenes at a level 100- to 1000-fold higher than the corresponding switch-deleted transgenes. To test whether the switch region of the natural IgH locus is also required for high level expression we have used homologous recombination to generate targeted recombinant hybridoma cell lines that lack the switch region sequences from the major intron of the mu gene. The expression pattern of these switch knock-out cell lines was compared with that of the parental cell line as well as to that of control recombinants using both steady-state mRNA level and nuclear run-on activity to assess heavy chain gene expression. In striking contrast with the results reported for transgenic animals, we have found only a minimal effect, if any, of deleting the switch element from the natural chromosomal location.
D000818:Animals; D017353:Gene Deletion; D005786:Gene Expression Regulation / Q000235:genetics*; D006825:Hybridomas; D007127:Immunoglobulin Constant Regions / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007075:Immunoglobulin M / Q000235:genetics*; D007134:Immunoglobulin Switch Region / Q000235:genetics*; D051379:Mice; D016297:Mutagenesis, Site-Directed / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis
D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D007075:Immunoglobulin M; D012333:RNA, Messenger