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Evaluation of macrophage-specific promoters using lentiviral delivery in mice.

Journal article
Authors Malin Levin
Ulf Lidberg
Pernilla Jirholt
Martin Adiels
Anna Wramstedt
K Gustafsson
D R Greaves
S Li
S Fazio
M F Linton
Sven-Olof Olofsson
Jan Borén
Inger Gjertsson
Published in Gene therapy
Volume 19
Issue 11
Pages 1041-7
ISSN 1476-5462
Publication year 2012
Published at Wallenberg Laboratory
Department of Mathematical Sciences
Institute of Medicine, Department of Rheumatology and Inflammation Research
Institute of Medicine, Department of Molecular and Clinical Medicine
Pages 1041-7
Language en
Links dx.doi.org/10.1038/gt.2011.195
Subject categories Clinical Medicine

Abstract

In gene therapy, tissue-specific promoters are useful tools to direct transgene expression and improve efficiency and safety. Macrophage-specific promoters (MSPs) have previously been published using different delivery systems. In this study, we evaluated five different MSPs fused with green fluorescent protein (GFP) to delineate the one with highest specificity using lentiviral delivery. We compared three variants of the CD68 promoter (full length, the 343-bp proximal part and the 150-bp proximal part) and two variants (in forward and reverse orientation) of a previously characterized synthetic promoter derived from elements of transcription factor genes. We transduced a number of cell lines and primary cells in vitro. In addition, hematopoietic stem cells were transduced with MSPs and transferred into lethally irradiated recipient mice. Fluorescence activated cell sorting analysis was performed to determine the GFP expression in different cell populations both in vitro and in vivo. We showed that MSPs can efficiently be used for lentiviral gene delivery and that the 150-bp proximal part of the CD68 promoter provides primarily macrophage-specific expression of GFP. We propose that this is the best currently available MSP to use for directing transgene expression to macrophage populations in vivo using lentiviral vectors.

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