A novel doxycycline-inducible system for the transgenic analysis of mammary gland biology
A novel doxycycline-inducible system for the transgenic analysis of mammary gland biology
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DOI：
10.1096/fj.01-0551com
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发表时间：
2002-03-01
期刊：
影响因子：
4.8
通讯作者：
Chodosh, LA
中科院分区：
文献类型：
--
作者：
Gunther, EJ;Belka, GK;Chodosh, LA
正常的发育事件，如青春期，怀孕，和奇偶性的影响乳腺肿瘤发生在人类和啮齿动物模型系统的易感性。不幸的是，依赖于乳腺特异性启动子来控制转基因表达的组成型转基因小鼠模型对于研究发育事件对乳腺癌风险的影响具有有限的效用，因为支配这些事件的激素信号也显著影响转基因表达水平。描述了一种新的转基因小鼠系统，其使用MMTV-LTR来驱动反向四环素依赖性反式激活因子rtTA的表达。将在乳腺上皮中表达rtTA的转基因小鼠与携带泰特操纵基因控制的转基因的报告细胞系杂交。我们测试的能力，空间，时间和定量控制报告基因的表达后，给予多西环素双转基因小鼠。使用该系统的转基因表达可以被快速诱导和去诱导，是高度乳腺特异性的，可以在广泛的表达水平范围内可重复地滴定，并且在未诱导状态下基本上是不可检测的。可以实现在整个乳腺上皮中的均质转基因表达。该系统允许将转基因表达限制在出生后乳腺发育的任何所需阶段。我们已经建立了一个乳腺特异性的，强力霉素诱导的转基因小鼠模型，用于研究乳腺发育对转基因介导的表型的影响。与已经描述的其它乳腺特异性转基因系统不同，该系统将青春期、妊娠、泌乳和退化期间乳腺上皮中的空间上均一的转基因表达与口服施用的廉价且广泛可用的诱导剂的使用相结合.该系统为乳腺生物学的转基因分析提供了新的机会。
Normal developmental events such as puberty, pregnancy, and parity influence the susceptibility of the mammary gland to tumorigenesis in both humans and rodent model systems. Unfortunately, constitutive transgenic mouse models that rely on mammary- specific promoters to control transgene expression have limited utility for studying the effect of developmental events on breast cancer risk since the hormonal signals governing these events also markedly influence transgene expression levels. A novel transgenic mouse system is described that uses the MMTV-LTR to drive expression of the reverse tetracycline-dependent transactivator rtTA. Transgenic mice expressing rtTA in the mammary epithelium were crossed with reporter lines bearing tet operator- controlled transgenes. We tested the ability to spatially, temporally, and quantitatively control reporter gene expression after administration of doxycycline to bi-transgenic mice. Transgene expression using this system can be rapidly induced and deinduced, is highly mammary specific, can be reproducibly titrated over a wide range of expression levels, and is essentially undetectable in the uninduced state. Homogeneous transgene expression throughout the mammary epithelium can be achieved. This system permits transgene expression to be restricted to any desired stage of postnatal mammary gland development. We have developed a mammary- specific, doxycycline- inducible transgenic mouse model for studying the effect of mammary gland development on transgene- mediated phenotypes. Unlike other mammary- specific, transgenic systems that have been described, this system combines spatially homogeneous transgene expression in the mammary epithelium during puberty, pregnancy, lactation, and involution with the use of an orally administered, inexpensive, and widely available inducing agent. This system offers new opportunities for the transgenic analysis of mammary gland biology in vivo.