Establishment of transgenic mice with IκBα low expression and the regulatory mechanism of the IκBα expression

LIU Qing-yan; XIA Ji-liang; XIAO Dong; PENG Bai-lu

Abstract：Objective In order to study the whole biological function of IκBαin vivo, we developed an IκBα low expression transgenic mice and analyze the regulatory mechanism of IκBα expression .Methods The targeting vector RV-IκBα-Loxp-Mut-Neo was constructed , then the linearized vector was injected into embryonic stem cells .South-ern blotting was used to screen the positive recombinant clones .Then the positive clone was microinjected into blastocysts to generate chimeric mice.Cross-breeding the chimeric with wild -type (C57BL/6N) mice generated heterozygous IκBα knockin mice .Homozygous IκBα knockin mice were cross -bred with Ella Cre mice to generate offspring , which interbred to generate homozygotes whose interval between two Loxp sites was deleted .The genetic type was identified with PCR . Western blot was used to detect expression of IκBα in various organs of transgenic mice .Results The IκBα knockin and IκBαknockout transgenic mice were established by PCR detection .The Western blot showed that the expression of IκBα protein in the spleen, thymus and lung of the homozygous IκBα knockin mice was lower than that in wild -type mice.A-part from the above organs , the mutant chromatid expression of IκBα protein in the skin , nasopharynx and stomach of the heterozygous IκBα knockin mice was also lower .Expression of IκBα protein was lower in most organs of IκBα knockout mice.Conclusion The transgenic mouse with low expression of IκBα was successfully established in the present study , and preliminary analysis of regulation of IκBα expression was implemented .