Construction and identification of conditional islet βcell DEPTOR knockout mice / 实用医学杂志

ABSTRACT

Objective To study the function of Deptor gene on the regulation of diabetes mellitus in suc-cessfully constructed and identified islet β-cell conditionally DEPTOR knockout mice model. Method By cross-breeding Deptorflox/floxmice with Cre mice expressed conditional specifically in pancreatic β-cell,Deptorflox/+Cre+/-mice were acquired and their genotypic identification was then performed. As the mice model of this study, Deptorflox/floxCre+/-mice were generated by crossing Deptorflox/+Cre+/-mice with Deptorflox/floxmice.Genotypic identifica-tion was performed by PCR at the age of 3 weeks. Tamoxifen was administered through intraperitoneal injection to induce the activation of the Cre recombination in islet beta cells of 8 weeks mice.Double immunofluorescence label-ing was then applied to identify the knockout effect of DEPTOR gene. Results Ten Islets Deptor knockout mice models were successfully acquired after 10-month cross-breeding. Validated genotype by PCR analysis were Deptorflox/floxCre+/- and double immunofluorescence labeling showed a significant difference between knockout mice and rodent controls. Conclusion Our study successfully constructs the islets conditionally Deptor deleted mice model by using Cre-loxp recombination system,providing a promising appliable animal model for study of dia-betes mellitus pathogenetic mechanism.