Temporal, spatial, and cell type-specific control of Cre-mediated DNA recombination in transgenic mice
Temporal, spatial, and cell type-specific control of Cre-mediated DNA recombination in transgenic mice
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DOI：
10.1038/15073
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发表时间：
1999-11-01
影响因子：
46.9
通讯作者：
Lee, WH
中科院分区：
文献类型：
--
作者：
Utomo, ARH;Nikitin, AY;Lee, WH
我们已经开发了一个通用系统的时间，空间和细胞类型特异性控制基因表达的小鼠，（1）整合四环素控制的基因表达和Cre-重组酶-loxP位点介导的基因失活的优势，（2）简化了动物杂交的方案，通过在一个单一的转基因的两个控制元件的组合。产生了两种转基因菌株，其中细胞类型特异性对照由视网膜母细胞瘤基因启动子或乳清酸性蛋白启动子提供。这两个启动子驱动反向四环素控制的反式激活因子（rtTA）的表达。rtTA诱导型启动子以顺式构型放置于rtTA转录单位，指导重组酶的表达。在与cActXstopXLacZ报告小鼠杂交的两个品系中，其具有由鸡β-肌动蛋白启动子驱动的loxP-转录终止/终止-loxP-LacZ盒，Cre-loxP介导的导致LacZ表达的DNA重组以时间、空间和细胞类型特异性方式被准确调控。这种方法可以应用于建立类似的小鼠品系，几乎任何启动子作为系统来控制各种细胞类型中的基因调控。
We have developed a universal system for temporal, spatial, and cell type-specific control of gene expression in mice that (1) integrates the advantages of tetracycline-controlled gene expression and Cre-recombinase-loxP site-mediated gene inactivation, and (2) simplifies schemes of animal crosses by combination of two control elements in a single transgene. Two transgenic strains were generated in which the cell type-specific control was provided by either the retinoblastoma gene promoter or the whey acidic protein promoter. Both promoters drive the expression of the reverse tetracycline-controlled transactivator (rtTA). Placed in cis configuration to the rtTA transcription unit, the rtTA-inducible promoter directs expression of ore recombinase. In both strains crossed with cActXstopXLacZ reporter mice, which have a loxP-stop of transcription/translation-loxP-LacZ cassette driven by chicken beta-actin promoter, Cre-loxP-mediated DNA recombination leading to LacZ expression was accurately regulated in a temporal, spatial, and cell type-specific manner. This approach can be applied to establishment of analogous mouse strains with virtually any promoter as systems to control gene regulation in a variety of cell types.