Nfkbia^tm1.1Pjc

Two well defined NFKB binding sites (M1 and M2) and 4 NFKB like binding sites (M3 - M6) in the promoter region were mutated, a FLAG tag was attached to exon 1, and a floxed neo cassette was inserted in intron 1 via homologous recombination. Cre mediated recombination removed the neo cassette. The locations of and mutations in the 6 binding sites are as follows: -115 - -93, a TCC motif changed to a CCG (M1); -143 - -122, a CCC motif changed to an AGT (M2); -227 - -205, a TCT motif changed to a CAC (M3); -292 - -271, a CC motif changed to a AA (M4); -319 - -297, a GGG motif changed to a TTT (M5); -387 - -365, a CCC motif changed to a GGG (M6). Chromatin immunoprecipitation analysis confirmed the loss of kappa B enhancer function in MEFs from homozygous mice. (J:163686)