Highly efficient gene transfer system using a laminin-DNA-apatite composite layer
Highly efficient gene transfer system using a laminin-DNA-apatite composite layer
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DOI：
10.1002/jgm.1425
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发表时间：
2010-02-01
影响因子：
3.5
通讯作者：
Ito, Atsuo
中科院分区：
文献类型：
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作者：
Oyane, Ayako;Tsurushima, Hideo;Ito, Atsuo
背景我们最近开发了一种使用层粘连蛋白-DNA-磷灰石复合层的安全有效的基因转移系统。本研究的目的是充分表征和优化层粘连蛋白-DNA-磷灰石复合层的基因转移效率，并证明在诱导细胞分化的复合层的可行性。方法层粘连蛋白-DNA-磷灰石复合层在各种条件下制备。使用荧光素酶和β-半乳糖苷酶基因表达测定系统评价所得复合层上的基因转移效率。结果层粘连蛋白-DNA-磷灰石复合层中的层粘连蛋白含量是提高基因转染效率的主要因素，而DNA含量不是影响基因转染效率的主要因素。层粘连蛋白在复合层中的细胞粘附性应该是基因转移效率提高的原因，因为在DNA-磷灰石复合层中没有细胞粘附性的白蛋白的固定对基因转移效率没有影响。结论本研究构建的基因转移系统具有安全、高效、可控的特点，可用于组织工程应用和基因转染芯片的制备。版权所有（C）2010约翰威利父子有限公司
Background We have recently developed a safe and efficient gene transfer system using a laminin-DNA-apatite composite layer. The objectives of the present study were to fully characterize and optimize the laminin-DNA-apatite composite layer in relation to the efficiency of gene transfer and to demonstrate the feasibility of the composite layer in the induction of cell differentiation.Methods The laminin-DNA-apatite composite layer was prepared under various conditions. The efficiency of gene transfer on the resulting composite layer was evaluated using luciferase and beta-galactosidase gene expression assay systems. A laminin-DNA-apatite composite layer, prepared under the optimized condition using a plasmid including cDNA of nerve growth factor (NGF), was then applied to the neuron-like differentiation of PC12 cells.Results The laminin content of the laminin-DNA-apatite composite layer was found to be a dominant factor improving the efficiency of gene transfer rather than the DNA content. The cell adhesion property of laminin in the composite layer should be responsible for the improvement in efficiency of gene transfer because the immobilization of albumin without the cell adhesion property in a DNA-apatite composite layer had no effect on the efficiency of gene transfer. A laminin-DNA-apatite composite layer, prepared under the optimized condition using a plasmid including cDNA of NGF, successfully induced the neuron-like differentiation of PC12 cells.Conclusions The present gene transfer system, with the potential to control cell differentiation and having features of safety and relatively high and controllable efficiency, would be a useful tool for tissue engineering applications and the production of transfection microarrays. Copyright (C) 2010 John Wiley & Sons, Ltd.