Ray M K, Fagan S P, Moldovan S, DeMayo F J, Brunicardi F C
Department of Surgery, Baylor College of Medicine, Houston, Texas 77030, USA.
Biochem Biophys Res Commun. 1998 Dec 9;253(1):65-9. doi: 10.1006/bbrc.1998.9714.
The rat insulin promoter (RIP) has been used to drive the expression of Cre recombinase (Cre) specifically in beta cells. Transient transfection was performed in the mouse insulinoma cell line, NIT-1, and control cell lines. RT-PCR was performed using total RNA from pancreas and other tissues of RIP-Cre transgenic mice. In addition, the efficiency and specificity of RIP were further analyzed by cross breeding the RIP-Cre transgenic mice with reporter mice bearing a beta-actin-loxP-CAT-loxP-lacZ transgene. In these mice, lacZ is expressed only after excision of the floxed-CAT gene by Cre-mediated recombination. Here, we present the data for beta cell-specific expression of lacZ in bigenic mice, as proof of concept in a mouse model for targeting beta cell-specific gene(s). The RIP-Cre transgenic mice will be used as a potential tool for targeting the excision of beta cell-specific gene(s) to study their role in islet cell physiology.
大鼠胰岛素启动子(RIP)已被用于驱动Cre重组酶(Cre)在β细胞中的特异性表达。在小鼠胰岛素瘤细胞系NIT-1和对照细胞系中进行了瞬时转染。使用来自RIP-Cre转基因小鼠胰腺和其他组织的总RNA进行RT-PCR。此外,通过将RIP-Cre转基因小鼠与携带β-肌动蛋白-loxP-CAT-loxP-lacZ转基因的报告小鼠杂交,进一步分析了RIP的效率和特异性。在这些小鼠中,只有在Cre介导的重组切除floxed-CAT基因后,lacZ才会表达。在这里,我们展示了双基因小鼠中lacZ在β细胞特异性表达的数据,作为在靶向β细胞特异性基因的小鼠模型中的概念验证。RIP-Cre转基因小鼠将作为一种潜在工具,用于靶向切除β细胞特异性基因,以研究它们在胰岛细胞生理学中的作用。