Influential factors on temperature-controlled gene expression using thermoresponsive polymeric gene carriers
Influential factors on temperature-controlled gene expression using thermoresponsive polymeric gene carriers
复制标题
热响应性聚合基因载体温控基因表达的影响因素
DOI：
10.1007/bf02481424
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发表时间：
2001
影响因子：
1.3
通讯作者：
T. Okano
中科院分区：
文献类型：
--
作者：
M. Yokoyama;M. Kurisawa;T. Okano
合成了3种温度响应性无规共聚物聚[N-异丙基丙烯酰胺（IPAAm）-co-2-（二甲氨基）乙基甲基丙烯酸酯（DMAEMA）-co-丁基甲基丙烯酸酯（BMA）]，并以β-半乳糖苷酶为标记基因，测定了其在不同孵育条件下对培养细胞的体外基因转染效率。当细胞孵育温度低于37°C时，这三种共聚物增加了基因表达。当温度降低时，转染效率的这种增加被认为是由于细胞中聚合物-DNA复合物的DNA释放增强。分析了复合物制备条件（电荷比和温度）和细胞孵育条件（用复合物孵育的时间、冷却温度、时间和时间段）的影响，以阐明该基因表达控制的机制，以及在降低细胞孵育温度时使基因表达的增加最大化。一种共聚物在温度降低到20°C持续3小时后显示基因表达增加了9倍。在分析复合物形成过程中温度对基因表达的影响时，认为紧密复合物形成有助于有效的细胞摄取和/或逃避酶促DNA降解。
Thermoresponsive random copolymers, poly[N-isopropylacrylamide (IPAAm)-co-2-(dimethylamino)ethyl methacrylate (DMAEMA)-co-butylmethacrylate (BMA)], were synthesized in three compositions, and their in vitro gene transfection efficiency to cultured cells at different incubation conditions was measured using β-galactosidase as a marker gene. These three copolymers increased gene expression when the cell incubation temperature was lowered below 37°C. This increase in transfection efficiency when temperature was lowered is considered to be due to enhanced DNA release from the polymer-DNA complex in cells. The influence of complex preparation conditions (charge ratio and temperature) and cell incubation conditions (incubation period with the complex, cooling temperature, timing, and period) was analyzed to elucidate the mechanism of this gene expression control as well as to maximize the increase in gene expression on lowering of the cell incubation temperature. One copolymer showed a nine-fold increase in gene expression on lowering of the temperature to 20°C for 3 h. In analyses of temperature effects in the complex formation procedure on the gene expression, tight complex formation was considered to contribute to efficient cell uptake and/or evasion from enzymatic DNA degradation.
通过在培养皿中与 β-半乳糖苷酶底物一起孵育细胞来进行 DNA 转染的简单测定。
影响因子：
2.7
作者：
Lim,K;Chae,CB
通讯作者：
Chae,CB
DOI：
10.1016/0006-291x(91)91366-k
发表时间：
1991-08-30
影响因子：
3.1
作者：
GAO, X;HUANG, L
通讯作者：
HUANG, L
DOI：
10.1073/pnas.84.21.7413
发表时间：
1987-11-01
影响因子：
11.1
作者：
FELGNER, PL;GADEK, TR;DANIELSEN, M
通讯作者：
DANIELSEN, M