The Terminator mouse is a diphtheria toxin-receptor knock-in mouse strain for rapid and efficient enrichment of desired cell lineages.

The Terminator mouse is a diphtheria toxin-receptor knock-in mouse strain for rapid and efficient enrichment of desired cell lineages.
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Terminator 小鼠是白喉毒素受体敲入小鼠品系,可快速有效地富集所需细胞谱系。

DOI:
10.1038/ki.2013.202
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发表时间:
2013-11
影响因子:
19.6
通讯作者:
--
中科院分区:
医学1区
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--
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生物医学研究通常需要特定细胞类型的原代培养,这是具有挑战性的,以可重复的方式获得高纯度。在这里,我们通过引入loxP位点两侧的白喉毒素受体来改造小鼠Rosa26位点。由此产生的菌株被戏称为终结者老鼠。这种方法导致白喉毒素受体在所有非cre表达细胞类型中表达,使这些细胞对白喉毒素暴露敏感。在终结者小鼠肾细胞原代培养中,超过99.99%的细胞在白喉毒素处理72小时内死亡。将终结者与Podocin-Cre(足细胞特异性)小鼠或Ggt-Cre(近端小管特异性)小鼠杂交后,白喉毒素处理杀死了不表达cre的细胞,但分别保留了足细胞和近端小管细胞,根据Western blotting和标记蛋白的免疫染色,原代培养物的纯度超过99%。因此,终结者小鼠可以成为一种有用的工具,可以选择性地和可重复地获得高数量和高纯度的低丰度细胞类型。
Biomedical research often requires primary cultures of specific cell types, which are challenging to obtain at high purity in a reproducible fashion. Here we engineered the murine Rosa26 locus by introducing the diphtheria toxin receptor flanked by loxP sites. The resultant strain was nicknamed the Terminator mouse. This approach results in diphtheria toxin receptor expression in all non-Cre expressing cell types, making these cells susceptible to diphtheria toxin exposure. In primary cultures of kidney cells derived from the Terminator mouse, over 99.99% of cells were dead within 72 hours of diphtheria toxin treatment. After crossing the Terminator with the Podocin-Cre (podocyte specific) mouse or the Ggt-Cre (proximal tubule specific) mouse, diphtheria toxin treatment killed non-Cre expressing cells but spared podocytes and proximal tubule cells, respectively, enriching the primary cultures to over 99% purity based on both Western blotting and immunostaining of marker proteins. Thus, the Terminator mouse can be a useful tool to selectively and reproducibly obtain even low abundant cell types at high quantity and purity.
DOI: 10.1371/journal.pone.0034907
发表时间: 2012
期刊: PloS one
影响因子: 3.7
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DOI: 10.1007/bf01233058
发表时间: 1991-07-01
期刊: SOMATIC CELL AND MOLECULAR GENETICS
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通讯作者: BROCKMAN, WW
DOI: 10.1002/mc.20036
发表时间: 2004-09-01
影响因子: 4.6
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