Division of Chemistry and Biological Chemistry, School of Physical and Mathematical Sciences, Nanyang Technological University, Singapore.
Biotechnol Genet Eng Rev. 2008;25:41-75. doi: 10.5661/bger-25-41.
Reporter gene assay is an invaluable tool for both biomedical and pharmaceutical researches to monitor cellular events associated with gene expression, regulation and signal transduction. On the basis of the alternations in reporter gene activities mediated by attaching response elements to these reporter genes, one sensitive, reliable and convenient assay can be provided to efficiently report the activation of particular messenger cascades and their effects on gene expression and regulations inside cells or living subjects. In this review, we introduce the current status of several commonly used reporter genes such as chloramphenicol acetyltransferase (CAT), alkaline phosphatase (AP), β-galactosidase (β-gal), luciferases, green fluorescent protein (GFP), and β-lactamase. Their applications in monitoring gene expression and regulations in vitro and in vivo will be summarized. With the development of advanced technology in gene expression and optical imaging modalities, reporter genes will become increasingly important in real-time detection of the gene expression at the single-cell level. This synergy will make it possible to understand the molecular basis of diseases, track the effectiveness of pharmaceuticals, monitor the response to therapies and evaluate the development process of new drugs.
报告基因检测是生物医学和药物研究中监测与基因表达、调控和信号转导相关的细胞事件的一种非常有价值的工具。通过将反应元件连接到这些报告基因上,可以介导报告基因活性的改变,从而提供一种灵敏、可靠和方便的检测方法,有效地报告特定信使级联的激活及其对细胞内或活体中基因表达和调控的影响。在这篇综述中,我们介绍了几种常用报告基因,如氯霉素乙酰转移酶(CAT)、碱性磷酸酶(AP)、β-半乳糖苷酶(β-gal)、荧光素酶、绿色荧光蛋白(GFP)和β-内酰胺酶的现状。总结了它们在体外和体内监测基因表达和调控中的应用。随着基因表达和光学成像模式的先进技术的发展,报告基因将在单细胞水平实时检测基因表达中变得越来越重要。这种协同作用将使我们有可能了解疾病的分子基础,跟踪药物的疗效,监测治疗反应,并评估新药的开发过程。