Guanidinium-grafted polyethylenimine: An efficient transfecting agent for mammalian cells
Guanidinium-grafted polyethylenimine: An efficient transfecting agent for mammalian cells
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DOI：
10.1016/j.ejpb.2007.09.008
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发表时间：
2008-03-01
影响因子：
4.9
通讯作者：
Chandra, Ramesh
中科院分区：
文献类型：
--
作者：
Nimesh, Surendra;Chandra, Ramesh
聚乙烯亚胺（PEI）是一种高效的多阳离子非病毒基因传递载体。其效率和细胞毒性取决于分子量，其中25 kda PEI效率最高，但伴随细胞毒性。在本研究中，探讨了通过胍侧基团的附着来提高基因传递效率和降低细胞毒性。假设胍定会导致聚合物的伯胺上电荷的离域，从而导致基因传递效率的提高以及细胞毒性的降低。用半硫酸o -甲基异脲对聚合物进行了胍化，并用红外光谱对其化学链进行了表征。用DLS法测定了胍基化PEI-DNA复合物的水动力直径。随后，以天然PEI为参考，将这些复合物用于DNA结合测定和ζ电位测量。进一步研究了胍基化PEI-DNA复合物对HEK 293细胞的基因传递效果。胍基化PEI-DNA复合物的水动力直径在176 ~ 548 nm之间。正如预期的那样，随着氮磷比的增加，zeta电位值增加。结果发现，在大多数测试的N/P比率下，与市售的转染剂脂质体和天然PEI本身相比，胍化PEI具有更高的转染效率。通过MTT比色法测定，与PEI聚合物相比，胍化PEI- dna复合物的毒性也大大降低。在制备的各种衍生物中，56%的gPEI在体外转染中被发现是最有效的。(C) 2007 Elsevier B.V.版权所有
Polyethylenimine (PEI) is one of the most efficient polycationic non-viral gene delivery vectors. Its efficiency and cytotoxicity depends on molecular weight, with the 25-kDa PEI being most efficient but accompanied with cytotoxicity. In the present study, enhancement in gene delivery efficiency along with reduction in cytotoxicity by attachment of guanidinium side group was explored. The hypothesis was that the guanidination would lead to the delocalization of charge present on primary amines of the polymer thereby leading to enhancement in gene delivery efficiency along with reduction in cytotoxicity. The polymer was guanidinated using O-methylisourea hemisulfate and the chemical linkage characterized by FTIR spectroscopy. The hydrodynamic diameter of guanidinated PEI-DNA complexes was determined using DLS. Subsequently, these complexes were used for DNA binding assay and zeta-potential measurements, taking native PEI as reference. Further, guanidinated PEI-DNA complexes were investigated for their gene delivery efficacy on HEK 293 cells. The hydrodynamic diameter of guanidinated PEI-DNA complexes was found to be in the range of 176-548 nm. As expected, the zeta potential values increased, on increasing the N/P ratios. It was found that guanidinated PEI had higher transfection efficiency at the majority of the N/P ratios tested as compared to commercially available transfecting agent lipofectin and native PEI itself. The toxicity of guanidinated PEI-DNA complexes was also reduced considerably in comparison to PEI polymer, as determined by MTT colorimetric assay. Out of the various derivatives prepared, gPEI 56% was found to be the most efficient in in vitro transfection. (C) 2007 Elsevier B.V. All rights reserved.