Generation and characterization of Ins1-cre-driver C57BL/6N for exclusive pancreatic beta cell-specific Cre-loxP recombination.
Generation and characterization of Ins1-cre-driver C57BL/6N for exclusive pancreatic beta cell-specific Cre-loxP recombination.
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DOI：
10.1538/expanim.63.183
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发表时间：
2014
影响因子：
2.4
通讯作者：
Yagami K
中科院分区：
文献类型：
--
作者：
Hasegawa Y;Daitoku Y;Mizuno S;Tanimoto Y;Mizuno-Iijima S;Matsuo M;Kajiwara N;Ema M;Oishi H;Miwa Y;Mekada K;Yoshiki A;Takahashi S;Sugiyama F;Yagami K
利用Cre/loxP系统介导的位点特异性重组研究基因在体内的功能。成功的有条件地敲除感兴趣的基因取决于Cre驱动的小鼠的可用性。我们培育并鉴定了胰腺β细胞特异性Cre驱动小鼠，用于糖尿病研究。将编码Cre的基因插入到小鼠Ins1的细菌人工染色体(BAC)第二外显子中。通过微量注射线性化BAC Ins1-cre建立了5只小鼠。转基因被整合在MafA和15号染色体上的端粒之间，在创始人之一BAC Ins1-CRE25中。为了研究Cre-loxP重组，BAC Ins1-cre25雄性与两个不同的Cre报告基因R26R和R26GRR雌性杂交。肉眼观察，Cre-loxP重组后的报告信号仅在两只F1小鼠的成年胰岛中检测到。免疫组织化学分析表明，Cre-loxP重组介导的报告信号在两个F1小鼠的胰岛细胞中与胰岛素共存，但不与胰高血糖素共存。此外，在胚胎13.5天，两个F1胎儿的胰岛中都已观察到Cre-loxP重组信号。最后，我们研究了Ins1的异位Cre-loxP重组，因为Ins2的同源基因除了在胰腺外，还在大脑中表达。然而，在两个F1小鼠的大脑中都没有Cre-loxP重组介导的报告信号。我们的数据表明，BACIns1-cre25小鼠是胰腺β细胞特异性Cre-loxP重组的有用的Cre驱动基因C57BL/6N，但与携带15号染色体上Flox基因的敲入小鼠杂交除外。
Cre/loxP system-mediated site-specific recombination is utilized to study gene function in vivo. Successful conditional knockout of genes of interest is dependent on the availability of Cre-driver mice. We produced and characterized pancreatic β cell-specific Cre-driver mice for use in diabetes mellitus research. The gene encoding Cre was inserted into the second exon of mouse Ins1 in a bacterial artificial chromosome (BAC). Five founder mice were produced by microinjection of linearized BAC Ins1-cre. The transgene was integrated between Mafa and the telomere on chromosome 15 in one of the founders, BAC Ins1-cre25. To investigate Cre-loxP recombination, BAC Ins1-cre25 males were crossed with two different Cre-reporters, R26R and R26GRR females. On gross observation, reporter signal after Cre-loxP recombination was detected exclusively in the adult pancreatic islets in both F1 mice. Immunohistological analysis indicated that Cre-loxP recombination-mediated reporter signal was colocalized with insulin in pancreatic islet cells of both F1 mice, but not with glucagon. Moreover, Cre-loxP recombination signal was already observed in the pancreatic islets at E13.5 in both F1 fetuses. Finally, we investigated ectopic Cre-loxP recombination for Ins1, because the ortholog Ins2 is also expressed in the brain, in addition to the pancreas. However, there was no Cre-loxP recombination-mediated reporter signal in the brain of both F1 mice. Our data suggest that BAC Ins1-cre25 mice are a useful Cre-driver C57BL/6N for pancreatic β cell-specific Cre-loxP recombination, except for crossing with knock-in mice carrying floxed gene on chromosome 15.
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影响因子：
29
作者：
Magnuson MA;Osipovich AB
通讯作者：
Osipovich AB
影响因子：
2.5
作者：
Mizuno, Seiya;Mizobuchi, Atsushi;Yagami, Ken-ichi
通讯作者：
Yagami, Ken-ichi
影响因子：
5.3
作者：
Cui, YX;Huang, L;Li, C
通讯作者：
Li, C
影响因子：
2.7
作者：
Gangalum, Rajendra K.;Jing, Zhe;Bhat, Suraj P.
通讯作者：
Bhat, Suraj P.
影响因子：
3.7
作者：
Katsumata T;Oishi H;Sekiguchi Y;Nagasaki H;Daassi D;Tai PH;Ema M;Kudo T;Takahashi S
通讯作者：
Takahashi S