| Project/Area Number |
06454262
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | Osaka University |
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Principal Investigator |
HAYASHI Norio Osaka University School of Medicine, Associate Professor, 医学部, 講師 (00144478)
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| Co-Investigator(Kenkyū-buntansha) |
HAGIWARA Hideki Osaka University Hospital Medical staff, 医学部・付属病院, 医員
MITA Eiji Osaka University Hospital Medical staff, 医学部・付属病院, 医員
TAKEHARA Tetsuo Osaka University Hospital Medical staff, 医学部・付属病院, 医員
KASAHARA Akinori Osaka University School of Medicine Assistant professor, 医学部, 助手 (70214286)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1995: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1994: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | in vitro gene transfer / in vivo gene transfer / asialoglycoprotein / cationic liposome / chloroquine / carrier-DNA ratio / gene therapy / selective gene delivery to the hepatocytes / アンチセンスDNA / ウイルス肝炎 / アシアログリコプロテイン / βガラクトシダーゼ / HcpG2 / 肝細胞 |
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| Research Abstract |
Asialoglycoprotein receptor mediated gene delivery is the most useful method for selective gene delivery to the hepatocytes. In vitro transfection of DNA to HepG2 cells, the transfection efficacy was highest at the carrier-DNA ratio of 480 : 1. Whne a lysosomal enzyme inhibitor, chloroquine was used for transfection into HepG2 cells, a 22-fold increase of efficacy was observed with addition of 50 mmol/L.The expression of transfered DNA-encoded products was observed in approximately 3% of HepG2 cells. When the carrier-^<32>P-labeled DNA complex was injected into the tail vessel of the mouse, most ^<32>P-labeled DNA were taken up by the liver. Microautoradiography of the live specimens after transfection of ^<35>S-labeled DNA indicated that the delivered DNA was located in the liver parenchymal cells.
When the efficacy of gene delivery was tested in vivo, chloroquine effectively increased the efficacy. Moreover, the combination of chloroquine and intraportal injection strongly enhanced the in vivo transfection with asialoglycoprotein receptor mediated gene delivery.
Thus, asialoglycoprotein receptor mediated gene transfer of anti-sense DNA is the selective gene delivery to the hepatocytes and may be the useful method of gene therapy for viral hepatitis.
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