Lin Weiying, Albanese Chris, Pestell Richard G, Lawrence David S
Department of Biochemistry, The Albert Einstein College of Medicine of Yeshiva University, Bronx, NY 10461, USA.
Chem Biol. 2002 Dec;9(12):1347-53. doi: 10.1016/s1074-5521(02)00288-0.
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 beta-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.
基于转基因的诱导表达系统为研究生物体生命周期中任何阶段任何基因的影响提供了可能。然而,单个基因的表达在时间和空间上(即细胞/组织水平)都是受到调控的。迄今为止设计的诱导基因表达系统无法对基因表达进行精细的空间控制。我们在此描述了一种光激活、蜕皮激素诱导的基因表达系统的构建及研究。我们构建了首个笼化蜕皮甾体的实例,在基于荧光素酶的基因表达系统中,它作为诱导剂实际上是无活性的。然而,在短暂光照下,笼化蜕皮甾体迅速转化为活性β - 蜕皮激素。笼化β - 蜕皮激素具有细胞渗透性,可在细胞内进行光解笼化,并且与点光源照明相结合,可用于在多细胞环境中驱动空间上离散的蛋白质表达。