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 language | English |
|---|---|
| Pages (from-to) | 145-148 |
| Number of pages | 4 |
| Journal | Neuroscience Letters |
| Volume | 324 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - May 17 2002 |
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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 journal › Article
}
TY - JOUR
T1 - Modification of a fiber protein in an adenovirus vector improves in vitro gene transfer efficiency to the mouse microglial cell line
AU - Omori, Nobuhiko
AU - Mizuguchi, Hiroyuki
AU - Ohsawa, Keiko
AU - Kohsaka, Shinichi
AU - Hayakawa, Takao
AU - Abe, Koji
AU - Shibasaki, Futoshi
PY - 2002/5/17
Y1 - 2002/5/17
N2 - 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.
AB - 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.
KW - Coxackievirus and adenovirus receptor
KW - Fiber mutant
KW - Microglia
KW - Recombinant adenovirus vector
KW - Reverse transcription-polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=0037123716&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037123716&partnerID=8YFLogxK
U2 - 10.1016/S0304-3940(02)00237-9
DO - 10.1016/S0304-3940(02)00237-9
M3 - Article
C2 - 11988348
AN - SCOPUS:0037123716
VL - 324
SP - 145
EP - 148
JO - Neuroscience Letters
JF - Neuroscience Letters
SN - 0304-3940
IS - 2
ER -