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The mouse Areg gene was replaced by human AREG coding sequence in B-hAREG MC38 cells. Human AREG is highly expressed in the suspensions of B-hAREG MC38 cells.
Gene targeting strategy for B-hAREG MC38 cells. The exogenous promoter and chimeric AREG coding sequence containing human extracellular domain, mouse transmembrane domain and cytoplasmic domain were inserted to replace part of murine exon 2 and all of exon 3 and exon 4. The insertion disrupts the endogenous murine Areg gene, resulting in a non-functional transcript.
B-hAREG MC38 cells were subcutaneously transplanted into C57BL/6N mice (n=5), and on 33 days post inoculation, tumor cells were harvested and assessed for human AREG expression by ELISA. As shown, human AREG was highly expressed in the suspensions of tumor cells. Therefore, B-hAREG MC38 cells can be used for in vivo efficacy studies of novel AREG therapeutics.