Ryan Amy, Scrable Heidi
University of Virginia School of Medicine, Charlottesville 22908-1392, USA.
Mol Imaging. 2004 Jan;3(1):33-42. doi: 10.1162/15353500200403193.
Reporter genes can monitor the status and activity of recombinant genomes in a diverse array of organisms, from bacteria and yeast to plants and animals. We have combined luciferase reporter genes with a conditional gene expression system based on regulatory elements from the lac operon of Escherichia coli to visualize the dynamics of gene expression in realtime in the living mouse. Using this technology, we have determined the rate of gene induction and repression, the level of target gene activity in response to different doses of inducer, and the schedule of induction during early embryogenesis of both the endogenous and the experimentally manipulated programs of mammalian gene expression associated with the HD/Hdh locus. The combination of in vivo imaging and lac regulation is a powerful tool for generating conditional transgenic mice that can be screened rapidly for optimal regulation and expression patterns, and for monitoring the induction and repression of regulated genes noninvasively in the living animal.
报告基因可监测从细菌、酵母到植物和动物等多种生物体中重组基因组的状态和活性。我们将荧光素酶报告基因与基于大肠杆菌乳糖操纵子调控元件的条件基因表达系统相结合,以实时观察活小鼠体内基因表达的动态变化。利用这项技术,我们已经确定了基因诱导和抑制的速率、响应不同剂量诱导剂时靶基因的活性水平,以及在哺乳动物基因表达的内源性和实验性操纵程序(与HD/Hdh基因座相关)的早期胚胎发育过程中的诱导时间表。体内成像和乳糖调控的结合是一种强大的工具,可用于生成条件转基因小鼠,这种小鼠能够快速筛选出最佳调控和表达模式,并用于在活体动物中无创监测调控基因的诱导和抑制。