Repetitive, non-invasive imaging of the dopamine D2 receptor as a reporter gene in living animals
Repetitive, non-invasive imaging of the dopamine D2 receptor as a reporter gene in living animals
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DOI：
10.1038/sj.gt.3300877
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发表时间：
1999-05-01
期刊：
影响因子：
5.1
通讯作者：
Herschman, HR
中科院分区：
文献类型：
--
作者：
MacLaren, DC;Gambhir, SS;Herschman, HR
报道基因（如β-半乳糖苷酶、氯霉素乙酰转移酶、绿色荧光蛋白、荧光素酶）在研究转基因动物的基因表达机制和开发用于基因治疗的基因递送系统中发挥作用。我们现在报告的程序，重复和非侵入性的图像报告基因表达的活动物与正电子发射断层扫描（PET）I使用多巴胺2型受体（D2 R）作为报告基因和3-（2 '[18 F]氟乙基）螺哌隆（FESP）作为报告探针。我们使用病毒递送系统来证明该PET报告基因/PET报告探针系统在体细胞基因转移后对报告基因表达成像的能力。在静脉注射携带D2 R报告基因的复制缺陷型腺病毒的小鼠中，肝脏[18 F]保留的体内PET测量与肝脏FESP保留、D2 R配体结合和D2 R mRNA的体外测量成比例。我们使用携带稳定转染的D2 R基因的肿瘤形成细胞来展示PET报告基因/PET报告探针系统在“组织”中的成像。表达转染的D2 R报告基因的肿瘤比对照肿瘤保留实质上更多的FESP。D2 R/FESP报告基因/报告探针系统应该是体内监测基因治疗载体和转基因表达的有价值的技术。
Reporter genes (eg beta-galactosiclase, chloramphenicol-acetyltransferase, green fluorescent protein, luciferase)play roles in investigating mechanisms of gene expression in transgenic animals and in developing gene delivery systems for gene therapy However, measuring expression of these reporter genes requires biopsy or death. We now report a procedure to image reporter gene expression repetitively and non-invasively in living animals with positron emission tomography (PET)I using the dopamine type 2 receptor (D2R) as a reporter gene and 3-(2'[18F]fluoroethyl)spiperone (FESP) as a reporter probe. We use a viral delivery system to demonstrate the ability of this PET reporter gene/PET reporter probe system to image repoter gene expression following somatic:gene transfer. in mice injected intravenously with replication-deficient adenovirus carrying a D2R reporter gene, PET in vivo measures of hepatic [18F] retention are proportional to in vitro measures of hepatic FESP retention, D2R ligand binding and D2R mRNA. We use tumor-forming cells carrying a stably transfected D2R gene to demonstrate imaging : of this PET reporter gene/PET reporter probe system in 'tissues'. Tumors expressing the transfected D2R reporter gene retain substantially more FESP than control tumors. The D2R/FESP reporter gene/reporter probe system should be a valuable technique to monitor, in vivo, expression from both gene therapy Vectors and transgenes.