1997 Fiscal Year Final Research Report Summary
Study for gene transfection and intracellular distribution of transfected DNA.
| Project/Area Number |
07558099
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Biophysics
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| Research Institution | Nagoya City University |
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Principal Investigator |
NAKANISHI Mamoru Nagoya City University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (90090472)
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| Co-Investigator(Kenkyū-buntansha) |
ITOH Hiroyasu Hamamatsu Photonics, Researcher, 研究員
FURUNO Tadahide Nagoya City University, Faculty of Pharmaceutical Sciences, Assistant professor, 薬学部, 講師 (80254308)
NANGO Mamoru Nagoya Institute of Technology, Associate professor, 工学部, 助教授 (90109893)
MIZUGUCHI Junichiro Tokyo Medical School, Professor, 教授 (20150188)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Cationic liposomes / Gene transfection / Cationic cholesterol / Zeta potential / Antisense DNA / Atomic force microscopy / Liposome / DNA complex / Plasmid DNA |
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| Research Abstract |
DNA-mediated transfection has become an important tool in modern biology. We have studied the use of the cationic liposomes for gene transfection. The results showed that the cationic liposomes containing a cationic cholesterol derivative, cholesteryl-3 beta -carboxyamido ethylenedimethylamine, were most effective among eight derivatives of cationic cholesterol. At the same time we measured zeta potential of the liposomes by laser Doppler spectroscopy. Values of zeta potential of the liposomes were well consistent with those of transfection efficiency of pSV2CAT into the cells. The results showed that zeta potential of cationic liposomes was one of the important factors which control gene transfection.
Next, we investigated that a novel cationic cholesterol derivative with a hydroxyethylamino head group. The transfection efficiency by the new cholesterol derivative was much higher than those by the cationic liposomes containing cationic derivatives with a dialkylamino head group described above. Further, the efficiency by the liposomes was not so much decreased in the presence of serum. This suggested that a novel cationic cholesterol derivative should be very promising in liposome-mediated gene transfection of plasmid and antisense DNA into target cells.
Further, atomic force microscopy (AFM) was used for studying gene transfection mediated by cationic liposomes which contain a cationic cholesterol derivative. AFM images showed that vesicles made of the liposome/DNA complex had various diameters depending on each cationic cholesterol derivative with a different spacer arm. The results showed that the diameter of the complex was well related to the transfection activity of plasmid DNA to cultured cell lines. From the results it was found that the vesicles with moderate diameters (from 0.4 to 1.4 mum) were most effective for gene transfection of plasmid DNA into the target cells.
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