Abstract
A series of thermoresponsive ternary random copolymers, poly[N-isopropylacrylamide (PIPAAm)-co-(dimethylamino)ethylmethacrylate (DMAEMA)-co-butylmethacrylate (BMA)], was synthesized and their in vitro gene transfection efficiency in cell culture was evaluated. A control copolymer containing 20 mol% DMAEMA units, IP-20D (mole ratio of IPAAm/DMAEMA/BMA=80/20/0 in feed, no BMA units) was inert in transfection. In contrast, copolymer IP-20D-10B (IPAAm/DMAEMA/BMA=70/20/10 in feed) effectively transfected plasmid DNA into COS-1 cell cultures even under small dosing conditions of 0.1 μg of plasmid DNA per well in a 96-well plate, suggesting that incorporation of the appropriate amount of hydrophobic unit is crucial to transfection efficiency. Gene expression was much more significant when transfected by the IP-20D-10B carrier in comparison with control homopolymer poly-DMAEMA, and almost equal to that of the highly competent lipid carrier, LipofectAMINE PLUS™. Furthermore, the transfection efficiency of IP-20D-10B is altered in a thermally responsive manner. By temporarily lowering the cell culture incubation temperature to 20°C in the posttransfection period, gene expression doubled over that for incubation temperature at 37°C. The DNA EtBr intercalation assay suggested that DNA affinity for IP-20D-10B is decreased by lowering incubation temperature, implying that the thermally regulated gene expression could provide more efficient DNA release from the polymeric carrier.
| Original language | English |
|---|---|
| Pages (from-to) | 343-355 |
| Number of pages | 13 |
| Journal | Journal of Controlled Release |
| Volume | 95 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - 2004 Mar 5 |
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Keywords
- Gene delivery
- Poly(N-isopropylacrylamide)
- Polymer carrier
- Thermoresponsive
- Transfection
ASJC Scopus subject areas
- Pharmaceutical Science
Cite this
Temperature-responsive polymeric carriers incorporating hydrophobic monomers for effective transfection in small doses. / Takeda, Naoya; Nakamura, Emiko; Yokoyama, Masayuki; Okano, Teruo.
In: Journal of Controlled Release, Vol. 95, No. 2, 05.03.2004, p. 343-355.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Temperature-responsive polymeric carriers incorporating hydrophobic monomers for effective transfection in small doses
AU - Takeda, Naoya
AU - Nakamura, Emiko
AU - Yokoyama, Masayuki
AU - Okano, Teruo
PY - 2004/3/5
Y1 - 2004/3/5
N2 - A series of thermoresponsive ternary random copolymers, poly[N-isopropylacrylamide (PIPAAm)-co-(dimethylamino)ethylmethacrylate (DMAEMA)-co-butylmethacrylate (BMA)], was synthesized and their in vitro gene transfection efficiency in cell culture was evaluated. A control copolymer containing 20 mol% DMAEMA units, IP-20D (mole ratio of IPAAm/DMAEMA/BMA=80/20/0 in feed, no BMA units) was inert in transfection. In contrast, copolymer IP-20D-10B (IPAAm/DMAEMA/BMA=70/20/10 in feed) effectively transfected plasmid DNA into COS-1 cell cultures even under small dosing conditions of 0.1 μg of plasmid DNA per well in a 96-well plate, suggesting that incorporation of the appropriate amount of hydrophobic unit is crucial to transfection efficiency. Gene expression was much more significant when transfected by the IP-20D-10B carrier in comparison with control homopolymer poly-DMAEMA, and almost equal to that of the highly competent lipid carrier, LipofectAMINE PLUS™. Furthermore, the transfection efficiency of IP-20D-10B is altered in a thermally responsive manner. By temporarily lowering the cell culture incubation temperature to 20°C in the posttransfection period, gene expression doubled over that for incubation temperature at 37°C. The DNA EtBr intercalation assay suggested that DNA affinity for IP-20D-10B is decreased by lowering incubation temperature, implying that the thermally regulated gene expression could provide more efficient DNA release from the polymeric carrier.
AB - A series of thermoresponsive ternary random copolymers, poly[N-isopropylacrylamide (PIPAAm)-co-(dimethylamino)ethylmethacrylate (DMAEMA)-co-butylmethacrylate (BMA)], was synthesized and their in vitro gene transfection efficiency in cell culture was evaluated. A control copolymer containing 20 mol% DMAEMA units, IP-20D (mole ratio of IPAAm/DMAEMA/BMA=80/20/0 in feed, no BMA units) was inert in transfection. In contrast, copolymer IP-20D-10B (IPAAm/DMAEMA/BMA=70/20/10 in feed) effectively transfected plasmid DNA into COS-1 cell cultures even under small dosing conditions of 0.1 μg of plasmid DNA per well in a 96-well plate, suggesting that incorporation of the appropriate amount of hydrophobic unit is crucial to transfection efficiency. Gene expression was much more significant when transfected by the IP-20D-10B carrier in comparison with control homopolymer poly-DMAEMA, and almost equal to that of the highly competent lipid carrier, LipofectAMINE PLUS™. Furthermore, the transfection efficiency of IP-20D-10B is altered in a thermally responsive manner. By temporarily lowering the cell culture incubation temperature to 20°C in the posttransfection period, gene expression doubled over that for incubation temperature at 37°C. The DNA EtBr intercalation assay suggested that DNA affinity for IP-20D-10B is decreased by lowering incubation temperature, implying that the thermally regulated gene expression could provide more efficient DNA release from the polymeric carrier.
KW - Gene delivery
KW - Poly(N-isopropylacrylamide)
KW - Polymer carrier
KW - Thermoresponsive
KW - Transfection
UR - http://www.scopus.com/inward/record.url?scp=1242338833&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=1242338833&partnerID=8YFLogxK
U2 - 10.1016/j.jconrel.2003.12.001
DO - 10.1016/j.jconrel.2003.12.001
M3 - Article
C2 - 14980782
AN - SCOPUS:1242338833
VL - 95
SP - 343
EP - 355
JO - Journal of Controlled Release
JF - Journal of Controlled Release
SN - 0168-3659
IS - 2
ER -