Podocyte-specific expression of tamoxifen-inducible Cre recombinase in mice
Podocyte-specific expression of tamoxifen-inducible Cre recombinase in mice
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DOI：
10.1093/ndt/gfq029
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发表时间：
2010-07-01
影响因子：
6.1
通讯作者：
Nakao, Kazuwa
中科院分区：
文献类型：
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作者：
Yokoi, Hideki;Kasahara, Masato;Nakao, Kazuwa
方法.为了在靶向足细胞的遗传实验中引入时间控制，我们构建了足细胞特异性启动子控制下的他莫昔芬诱导的Cre重组酶（CreER（T2））转基因小鼠，2.5 kb片段的人podocin（NPHS 2）基因。通过将NPHS 2-CreER（T2）与ROSA 26报告基因（R26 R）小鼠杂交来检查Cre活性的特异性和效率，在所述R26 R小鼠中，终止终止盒已被置于β-半乳糖苷酶基因的上游。4周龄的双突变小鼠（NPHS 2-CreER（T2）/R26 R）连续3天腹腔注射0.5 mg 4-羟基他莫昔芬（4-OHT）。用4-OHT处理的NPHS 2-CreER（T2）/R26 R在肾小球中85%的足细胞中特异性表达β-半乳糖苷酶。Cre重组酶mRNA的表达主要局限于肾脏，尤其是肾小球。总之，我们已经成功地产生足细胞特异性诱导Cre转基因小鼠他莫昔芬管理。这些小鼠使我们能够在出生后破坏足细胞中的基因。
Methods. To introduce a temporal control in the genetic experiments targeting the podocyte, we constructed tamoxifen-inducible Cre recombinase (CreER(T2)) transgenic mice under the control of podocyte-specific promoter, 2.5-kb fragment of the human podocin (NPHS2) gene. The specificity and efficiency of Cre activity were examined by crossing NPHS2-CreER(T2) with the ROSA26 reporter (R26R) mouse in which a floxed-stop cassette has been placed upstream of the beta-galactosidase gene. Four-week-old double-mutant mice (NPHS2-CreER(T2)/R26R) were intraperitoneally administered with 0.5 mg of 4-hydroxytamoxifen (4-OHT) for three consecutive days.Results. NPHS2-CreER(T2)/R26R treated with 4-OHT expressed beta-galactosidase specifically in 85% of the podocytes in glomeruli. Expression of Cre recombinase mRNA was mostly restricted to the kidney, especially in glomeruli.Conclusions. In conclusion, we have successfully generated podocyte-specific inducible Cre transgenic mice by tamoxifen administration. These mice allow us to disrupt the genes specifically in the podocytes after birth.