Evaluation of macrophage-specific promoters using lentiviral delivery in mice

M. C. Levin, U. Lidberg, P. Jirholt, M. Adiels, A. Wramstedt, K. Gustafsson, D. R. Greaves, S. Li, Sergio Fazio, M. F. Linton, S. O. Olofsson, J. Borén, I. Gjertsson

Research output: Contribution to journalArticle

12 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1041-1047
Number of pages7
JournalGene Therapy
Volume19
Issue number11
DOIs
StatePublished - Nov 2012
Externally publishedYes

Fingerprint

Macrophages
Green Fluorescent Proteins
Transgenes
Hematopoietic Stem Cells
Genetic Therapy
Population
Genes
Flow Cytometry
Transcription Factors
Safety
Cell Line
In Vitro Techniques

Keywords

  • Lentivirus
  • Macrophage
  • Promoter

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Levin, M. C., Lidberg, U., Jirholt, P., Adiels, M., Wramstedt, A., Gustafsson, K., ... Gjertsson, I. (2012). Evaluation of macrophage-specific promoters using lentiviral delivery in mice. Gene Therapy, 19(11), 1041-1047. https://doi.org/10.1038/gt.2011.195

Evaluation of macrophage-specific promoters using lentiviral delivery in mice. / Levin, M. C.; Lidberg, U.; Jirholt, P.; Adiels, M.; Wramstedt, A.; Gustafsson, K.; Greaves, D. R.; Li, S.; Fazio, Sergio; Linton, M. F.; Olofsson, S. O.; Borén, J.; Gjertsson, I.

In: Gene Therapy, Vol. 19, No. 11, 11.2012, p. 1041-1047.

Research output: Contribution to journalArticle

Levin, MC, Lidberg, U, Jirholt, P, Adiels, M, Wramstedt, A, Gustafsson, K, Greaves, DR, Li, S, Fazio, S, Linton, MF, Olofsson, SO, Borén, J & Gjertsson, I 2012, 'Evaluation of macrophage-specific promoters using lentiviral delivery in mice', Gene Therapy, vol. 19, no. 11, pp. 1041-1047. https://doi.org/10.1038/gt.2011.195
Levin MC, Lidberg U, Jirholt P, Adiels M, Wramstedt A, Gustafsson K et al. Evaluation of macrophage-specific promoters using lentiviral delivery in mice. Gene Therapy. 2012 Nov;19(11):1041-1047. https://doi.org/10.1038/gt.2011.195
Levin, M. C. ; Lidberg, U. ; Jirholt, P. ; Adiels, M. ; Wramstedt, A. ; Gustafsson, K. ; Greaves, D. R. ; Li, S. ; Fazio, Sergio ; Linton, M. F. ; Olofsson, S. O. ; Borén, J. ; Gjertsson, I. / Evaluation of macrophage-specific promoters using lentiviral delivery in mice. In: Gene Therapy. 2012 ; Vol. 19, No. 11. pp. 1041-1047.
@article{6e83c185315f4a27aecc51ce7c5d32c1,
title = "Evaluation of macrophage-specific promoters using lentiviral delivery in mice",
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.",
keywords = "Lentivirus, Macrophage, Promoter",
author = "Levin, {M. C.} and U. Lidberg and P. Jirholt and M. Adiels and A. Wramstedt and K. Gustafsson and Greaves, {D. R.} and S. Li and Sergio Fazio and Linton, {M. F.} and Olofsson, {S. O.} and J. Bor{\'e}n and I. Gjertsson",
year = "2012",
month = "11",
doi = "10.1038/gt.2011.195",
language = "English (US)",
volume = "19",
pages = "1041--1047",
journal = "Gene Therapy",
issn = "0969-7128",
publisher = "Nature Publishing Group",
number = "11",

}

TY - JOUR

T1 - Evaluation of macrophage-specific promoters using lentiviral delivery in mice

AU - Levin, M. C.

AU - Lidberg, U.

AU - Jirholt, P.

AU - Adiels, M.

AU - Wramstedt, A.

AU - Gustafsson, K.

AU - Greaves, D. R.

AU - Li, S.

AU - Fazio, Sergio

AU - Linton, M. F.

AU - Olofsson, S. O.

AU - Borén, J.

AU - Gjertsson, I.

PY - 2012/11

Y1 - 2012/11

N2 - 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.

AB - 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.

KW - Lentivirus

KW - Macrophage

KW - Promoter

UR - http://www.scopus.com/inward/record.url?scp=84869092003&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84869092003&partnerID=8YFLogxK

U2 - 10.1038/gt.2011.195

DO - 10.1038/gt.2011.195

M3 - Article

C2 - 22130447

AN - SCOPUS:84869092003

VL - 19

SP - 1041

EP - 1047

JO - Gene Therapy

JF - Gene Therapy

SN - 0969-7128

IS - 11

ER -