Establishment of <italic>Muc1</italic> Gene Knockout Mouse Model

Abstract

MUCIN1 (MUC1), a member of the mucin family anchored to the apical surface of the epithelial cells, is responsible for immune inflammation and tumorigenesis. A Muc1 knockout mouse model was established for exploring the biological functions and studying mechanisms of MUC1 in tumorigenesis and metastasis. Mouse genomic DNA sequence of the Muc1 gene was obtained using bioinformatics methods. Muc1 gene knockout vector Muc1 -ABRLFn- pBR322 was constructed with exon 2 and exon 3 flanked by two loxP sites. Muc1 knockout vector was transferred into the embryonic stem cells by electroporation and G418 and Ganciclovoir resistant clones were screened. Four correctly homologous clones were identified by PCR, one of which was microinjected into C57BL/6J mouse blastocysts to obtain chimera mice. Sixteen mice with a chimera rate of over 50% were acquired out of 32 bred by 13 receptors. Chimeric mice were mated with C57BL/6J to obtain 11 heterozygous mice (10 males and one female). Heterozygous mice were backcrossed and then mated with EIIa-Cre mice. Thus Muc1 gene knockout mice model was successfully created and there was no embryonic lethality in homologous mutant mice. Preliminary observation of phenotype revealed no abnormalities in structures in relevant tissues and organs of Muc1 knockout mice.