Visualizing gene expression in living mammals using a bioluminescent reporter
Visualizing gene expression in living mammals using a bioluminescent reporter
复制标题
DOI：
10.1111/j.1751-1097.1997.tb03184.x
复制
发表时间：
1997-10-01
影响因子：
3.3
通讯作者：
Benaron, DA
中科院分区：
文献类型：
--
作者：
Contag, CH;Spilman, SD;Benaron, DA
基因表达的控制通常涉及一系列相互交织的调控过程。由于调控途径的信息可能会丢失在体外的分析中，我们使用生物发光监测在活体哺乳动物中的基因表达，病毒启动子融合到萤火虫荧光素酶作为转基因小鼠允许外部的监测基因的表层和深层组织中的表达，在体内的生物发光既可用增强或冷却的电荷耦合设备相机检测到，并可在局部和系统递送底物后检测到；体内控制人类免疫缺陷病毒的启动子被证明，作为基于DNA的治疗和疫苗的模型，在体内转染荧光素酶表达载体(SV-40启动子和增强子控制表达)。我们的结论是，现在可以使用荧光素酶报告程序对活体哺乳动物的基因调控、DNA递送和表达进行非侵入性监测，因此，对人类发育和疾病的活体动物模型中的基因表达进行实时、非侵入性研究是可能的。
Control of gene expression often involves an interwoven set of regulatory processes. As information regarding regulatory pathways may be lost in ex vivo analyses, we used bioluminescence to monitor gene expression in living mammals, Viral promoters fused to firefly luciferase as transgenes in mice allowed external monitoring of gene expression both superficially and in deep tissues, In vivo bioluminescence was detectable using either intensified or cooled charge-coupled device cameras, and could be detected following both topical and systemic delivery of substrate, In vivo control of the promoter from the human immunodeficiency virus was demonstrated, As a model for DNA-based therapies and vaccines, in vivo transfection of a luciferase expression vector (SV-40 promoter and enhancer controlling expression) was detected. We conclude that gene regulation, DNA delivery and expression can now be noninvasively monitored in living mammals using a luciferase reporter, Thus, real-time, noninvasive study of gene expression in living animal models for human development and disease is possible.