Spatially discrete, light-driven protein expression
Spatially discrete, light-driven protein expression
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DOI：
10.1016/s1074-5521(02)00288-0
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发表时间：
2002-12-01
影响因子：
--
通讯作者：
Lawrence, DS
中科院分区：
文献类型：
--
作者：
Lin, WY;Albanese, C;Lawrence, DS
基于转基因的诱导表达系统提供了研究生物体生命周期中任何基因在任何时间点的影响的潜力。然而，单个基因的表达在时间和空间上都是一致的（即，细胞/组织）调节。迄今为止设计的诱导型基因表达系统并不能对基因表达提供精细的空间控制。我们在此描述了一个光激活的蜕皮激素诱导的基因表达系统的创建和研究。我们已经构建了第一个笼状蜕皮甾体的例子，这是几乎无活性的诱导剂在基于酶的基因表达系统。然而，在暴露于短暂的光照下时，笼状蜕皮类固醇迅速转化为活性β-蜕皮激素。笼状β-蜕皮激素具有细胞渗透性，可以在细胞内进行光捕获，并且与点照明相结合，可用于在多细胞环境中驱动空间离散的蛋白质表达。
Transgene-based inducible expression systems offer the potential to study the influence of any gene at any point during an organism's lifetime. However, the expression of individual genes is both temporally and spatially (i.e., cell/tissue)-regulated. The inducible gene expression systems devised to date do not offer fine spatial control over gene expression. We describe herein the creation and study of a light-activatable, ecdysone-inducible gene expression system. We have constructed the first example of a caged ecdysteroid, which is virtually inactive as an inducing agent in a luciferase-based gene expression system. However, upon exposure to brief illumination, the caged ecdysteroid is rapidly converted into active beta-ecdysone. Caged p-ecdysone is cell permeable, can be intracellularly photouncaged, and, in combination with spot illumination, can be used to drive spatially discrete protein expression in a multicellular setting.