Guo Z S, Wang L H, Eisensmith R C, Woo S L
Department of Cell Biology, Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030, USA.
Gene Ther. 1996 Sep;3(9):802-10.
Transgene expression in studies of both gene function and gene therapy may be assisted considerably through the use of transcriptional regulatory elements which permit high-level, and/or tissue-specific gene expression. We have therefore evaluated the transcriptional activities of a series of viral and cellular enhancer/promoter elements, both in vitro and in vivo. The five enhancer/promoter elements showing either high-level or hepatocyte-specific expression following transient transfection into hepatoma cells were incorporated into recombinant adenoviruses expressing human alpha 1-antitrypsin (hAAT) for in vivo studies in the liver of immunodeficient and immunocompetent mice. The human elongation factor 1 alpha gene promoter produced 2 mg/ml serum level of hAAT, which is physiologic in humans and will be therapeutic for patients with AAT deficiency. This and all other enhancer/promoters except that of the CMV-IE gene yielded persistent hAAT expression in SCID mice. These findings demonstrate that adenovirus vectors provide an effective system for studies designed to evaluate enhancer/promoter activities in vivo. Several of the enhancer/promoters examined in this study will have significant utility in adenovirus-mediated gene therapy for alpha 1-antitrypsin deficiency and other genetic disorders.
在基因功能研究和基因治疗中，通过使用能够实现高水平和/或组织特异性基因表达的转录调控元件，转基因表达可得到显著促进。因此，我们在体外和体内评估了一系列病毒和细胞增强子/启动子元件的转录活性。将在瞬时转染入肝癌细胞后表现出高水平或肝细胞特异性表达的五个增强子/启动子元件，整合到表达人α1-抗胰蛋白酶（hAAT）的重组腺病毒中，用于在免疫缺陷和免疫健全小鼠肝脏中的体内研究。人延伸因子1α基因启动子产生的hAAT血清水平为2mg/ml，这在人体内是生理水平，对α1-抗胰蛋白酶缺乏症患者具有治疗作用。除CMV-IE基因的增强子/启动子外，该增强子/启动子以及所有其他增强子/启动子在SCID小鼠中均产生了持续的hAAT表达。这些发现表明，腺病毒载体为旨在评估体内增强子/启动子活性的研究提供了一个有效的系统。本研究中检测的几种增强子/启动子在腺病毒介导的α1-抗胰蛋白酶缺乏症和其他遗传疾病的基因治疗中将具有重要用途。
