增强子
生物
病毒载体
报告基因
人巨细胞病毒
转导(生物物理学)
转基因
基因表达
分子生物学
绿色荧光蛋白
遗传增强
基因
遗传学
生物化学
重组DNA
作者
Ina Gruh,Stephanie Wunderlich,M Winkler,Kristin Schwanke,Jennifer Heinke,Ulrike Blömer,Arjang Ruhparwar,B Rohde,Ren‐Ke Li,Axel Haverich,Ulrich Martin
摘要
Abstract Background Lentiviral vectors are attractive delivery tools for gene therapy, especially in terminally differentiated target cells. While restriction of gene expression to specific cell populations is of particular importance, highly efficient cell‐type‐specific gene expression after viral gene transfer so far has been hampered by low levels of transgene expression. Methods Addressing this problem, we have integrated the human cytomegalovirus (CMV) immediate‐early enhancer into an ‘advanced’ generation lentiviral vector. Expression cassettes with the reporter gene green fluorescent protein (GFP), combined with the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) under control of a ubiquitous phosphoglycerate kinase (mouse PGK), cardiomyocyte‐ (human atrial natriuretic factor (ANF), human ventricular myosin light chain (MLC2v)), or type II alveolar epithelial cell (AT‐2)‐specific human surfactant protein C (SP‐C) promoter, were introduced. As insertion of an enhancing element can interfere with the promoter's specificity, expression levels conferred by our enhancer/promoter constructs were evaluated in target and non‐target cells. Results Transduction of target cells with human CMV enhancer containing lentiviral vectors resulted in a multiple‐log increase in GFP expression compared to corresponding vectors lacking the human CMV enhancer. In the case of the ANF, the MLC2v, and the SP‐C promoters, tissue‐specific reporter gene expression in cardiomyocytes and in lung AT‐2 cells was maintained, as expression in non‐target cells increased only up to 7‐fold. Conclusions The results of this study indicate that lentiviral vectors with the human CMV enhancer conferring efficient cell‐type‐specific gene expression may be useful tools for gene therapy purposes or cell tracing, e.g. to analyze stem cell differentiation in transplantation and co‐culture settings. Copyright © 2007 John Wiley & Sons, Ltd.
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