2001 Fiscal Year Final Research Report Summary
Gene Delivery System Using Thermo-responsive Polymers
| Project/Area Number |
11650913
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
高分子合成
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| Research Institution | Tokyo Woman's Medical University |
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Principal Investigator |
YOKOYAMA Masayuki School of Medicine, Tokyo Woman's Medical University, Asistant Professor, 医学部, 助教授 (20220577)
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| Project Period (FY) |
1999 – 2001
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| Keywords | Thermo-response / Synthetic Polymer / Gene / DNA / Delivery / Synthetic Vector / Gene expression / Complex |
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| Research Abstract |
A thermo-responsive copolymer, poly(N-isopropylacrylamjde (IPAAm)-co-2-(dimethylamino)ethyl methacrylate (DMAEMA)-co-butylmethacrylate (BMA)) was synthesized, and its in vitro gene transfection efficiency at different incubation temperatures was evaluated. A copolymer containing 8 mol% of DMAEMA and 11 mol% of BMA (P(IP-8DA-11BM)) had a lower critical solution temperature (LCST) at 21℃, therefore, the copolymer was insoluble above 21℃ and soluble below 21℃. This copolymer was complexed with plasmid DNA. Transfection efficiency of polymer-plasmid complexes was evaluated in COS-1 cells using pCMV-βgal plasmid, encoding for β-galactosidase as a reporter gene. Transfection efficiency of PDMAEMA homopolymer incubated at 37℃ for 48h was higher than that incubated at 20℃ for 3h and 37℃ for 45h. In contrast, transfection efficiency of P(IP-8DA-11BM) incubated at 20℃ for 3h and 37℃ for 45h was much higher than that incubated at 37℃ for 48h. This gene expression enhancement was increased to 8.6-fold by optimizing incubation period of the complex with cells. Such increased transfection efficiency by lowering temperature is considered to be due to appropriate formation/dissociation control of P(IP-8DA-11BM)-DNA complexes.
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