Honkakoski P, Jääskeläinen I, Kortelahti M, Urtti A
Department of Pharmaceutics, University of Kuopio, Finland.
Pharm Res. 2001 Feb;18(2):146-50. doi: 10.1023/a:1011068015301.
To develop and characterize a new drug-regulated gene expression system based on the nuclear receptor constitutive androstane receptor (CAR).
Both transient and stable transfection into HEK293 cells of luciferase plasmids under the control of either drug- and steroid-responsive nuclear receptor CAR or the tetracycline-sensitive transactivator tTA were used in development of stable cell lines.
A stable first-generation cell line that expresses luciferase gene under the control of nuclear receptor CAR was developed. The luciferase expression in CAR-producing cells could be suppressed by androstanes and reactivated by structurally unrelated drugs chlorpromazine, metyrapone, phenobarbital, and clotrimazole. The kinetics of luciferase expression in CAR-producing cells and the tTA system were comparable. The overall regulation of CAR system was improved by modifications to the DNA binding domain and site.
Because of its wide ligand selectivity and transferable ligand binding domain, CAR expands the repertoire of regulated gene expression systems.
基于核受体组成型雄甾烷受体（CAR）开发并表征一种新型药物调控基因表达系统。
在稳定细胞系的开发中，将受药物和类固醇响应性核受体CAR或四环素敏感性反式激活因子tTA控制的荧光素酶质粒瞬时和稳定转染至HEK293细胞中。
开发出了一种在核受体CAR控制下表达荧光素酶基因的稳定第一代细胞系。产生CAR的细胞中的荧光素酶表达可被雄甾烷抑制，并被结构不相关的药物氯丙嗪、甲吡酮、苯巴比妥和克霉唑重新激活。产生CAR的细胞中荧光素酶表达的动力学与tTA系统相当。通过对DNA结合结构域和位点的修饰，改善了CAR系统的整体调控。
由于其广泛的配体选择性和可转移的配体结合结构域，CAR扩展了受调控基因表达系统的种类。
