These authors contributed equally to this work.
Human CMV immediate-early enhancer: a useful tool to enhance cell-type-specific expression from lentiviral vectors
Version of Record online: 19 NOV 2007
Copyright © 2007 John Wiley & Sons, Ltd.
The Journal of Gene Medicine
Volume 10, Issue 1, pages 21–32, January 2008
How to Cite
Gruh, I., Wunderlich, S., Winkler, M., Schwanke, K., Heinke, J., Blömer, U., Ruhparwar, A., Rohde, B., Li, R.-K., Haverich, A. and Martin, U. (2008), Human CMV immediate-early enhancer: a useful tool to enhance cell-type-specific expression from lentiviral vectors. J. Gene Med., 10: 21–32. doi: 10.1002/jgm.1122
- Issue online: 17 DEC 2007
- Version of Record online: 19 NOV 2007
- Manuscript Accepted: 18 SEP 2007
- Manuscript Revised: 9 AUG 2007
- Manuscript Received: 6 MAR 2007
- HILF program of the Hannover Medical School, the Niedersachsen Stiftung/CORTISS and by the Lower Saxony Ministry for Education and Science to fund joint Lower Saxony-Israel research projects
- lentiviral vector;
- gene transfer;
- tissue-specific expression;
- CMV enhancer
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.
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.
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.
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.