Modification of a fiber protein in an adenovirus vector improves in vitro gene transfer efficiency to the mouse microglial cell line

Nobuhiko Omori, Hiroyuki Mizuguchi, Keiko Ohsawa, Shinichi Kohsaka, Takao Hayakawa, Koji Abe, Futoshi Shibasaki

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

In microglia, it is difficult to introduce exogenous genes of interest even by recombinant adenovirus vectors (Ad) which can infect with high efficiency only to the cells expressing coxackievirus and adenovirus receptors (CAR). We found a lack of CAR expression in primary cultured murine microglia (PCMG) and its immortalized cell line MG5 by reverse transcription-polymerase chain reaction. In order to improve the efficiency of gene transfer, we generated a novel Ad (Ad-RGD) by an incorporation of the Arg-Gly-Asp motif (RGD) containing peptide in the HI loop of the viral fiber knob domain, which enables the virus to contact target cells through αV integrins which are known to be ubiquitously expressed on the surface of mammalian cells. Ad-RGD showed a remarkable improvement (13-18-fold) in the delivery of Escherichia coli LacZ gene in MG5 cells and a moderate increase in PCMG cells under the treatment with granulocyte-macrophage colony stimulating factor. These results suggest that Ad-RGD may be a potent tool for the delivery of genes to microglia activated by optimum stimulation, and thus analyzing the function of microglia with utilization of MG5 and PCMG cells.

Original languageEnglish
Pages (from-to)145-148
Number of pages4
JournalNeuroscience Letters
Volume324
Issue number2
DOIs
Publication statusPublished - May 17 2002

Fingerprint

Microglia
Cell Line
Genes
Peptide PHI
Lac Operon
Granulocyte-Macrophage Colony-Stimulating Factor
Adenoviridae
Integrins
Reverse Transcription
Adenovirus hexon capsid protein
In Vitro Techniques
Escherichia coli
Viruses
Polymerase Chain Reaction

Keywords

  • Coxackievirus and adenovirus receptor
  • Fiber mutant
  • Microglia
  • Recombinant adenovirus vector
  • Reverse transcription-polymerase chain reaction

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Modification of a fiber protein in an adenovirus vector improves in vitro gene transfer efficiency to the mouse microglial cell line. / Omori, Nobuhiko; Mizuguchi, Hiroyuki; Ohsawa, Keiko; Kohsaka, Shinichi; Hayakawa, Takao; Abe, Koji; Shibasaki, Futoshi.

In: Neuroscience Letters, Vol. 324, No. 2, 17.05.2002, p. 145-148.

Research output: Contribution to journalArticle

Omori, Nobuhiko ; Mizuguchi, Hiroyuki ; Ohsawa, Keiko ; Kohsaka, Shinichi ; Hayakawa, Takao ; Abe, Koji ; Shibasaki, Futoshi. / Modification of a fiber protein in an adenovirus vector improves in vitro gene transfer efficiency to the mouse microglial cell line. In: Neuroscience Letters. 2002 ; Vol. 324, No. 2. pp. 145-148.
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