Mechanisms involved in gene electrotransfer using high- and low-voltage pulses - An in vitro study
Mechanisms involved in gene electrotransfer using high- and low-voltage pulses - An in vitro study
复制标题
DOI：
10.1016/j.bioelechem.2008.09.002
复制
发表时间：
2009-02-01
影响因子：
5
通讯作者：
Pavlin, Mojca
中科院分区：
文献类型：
--
作者：
Kanduser, Masa;Miklavcic, Damijan;Pavlin, Mojca
基因电转移是一种已建立的基因递送方法，其使用高压脉冲来增加细胞膜的渗透性，从而能够转移基因。目前的研究主要集中在提高体内转染效率，而基因电转移的机制还不完全清楚，本文分析了高压和低压脉冲联合作用下基因电转移的机制。我们将HV和LV脉冲的不同组合应用于CHO细胞并测定转染效率。我们得到，短HV脉冲单独足以将DNA递送到细胞中以获得最佳质粒浓度，并且LV脉冲不增加转染效率，与体内报告的研究相反。然而，对于次优质粒浓度，HV和LV脉冲的组合增加了转染率。我们的研究结果表明，低电压脉冲增加转染生病的条件下，质粒浓度低，通常在体内的DNA的流动性是有限的细胞外基质。LV脉冲提供了额外的电泳力，其将DNA拖向细胞膜并因此增加转染效率，而对于足够高浓度的质粒（通常在体外使用），电泳LV脉冲不具有重要作用。(C)2008 Elsevier B.V.保留所有权利。
Gene electrotransfer is an established method for gene delivery which uses high-voltage pulses to increase permeability of cell membrane and thus enables transfer of genes. Currently, majority of research is focused on improving in vivo transfection efficiency, while mechanisms involved in gene electrotransfer are not completely understood.In this paper we analyze the mechanisms of gene electrotransfer by using combinations of high-voltage (HV) and low-voltage pulses (LV) in vitro. We applied different combinations of HV and LV Pulses to CHO cells and determined the transfection efficiency. We obtained that short HV pulses alone were sufficient to deliver DNA into cells for optimal plasmid concentrations and that LV pulse did not increase transfection efficiency, in contrast to reported studies in vivo. However, for sub-optimal plasmid concentrations combining HV and LV pulses increased transfection rate. Our results Suggest that low-voltage pulses increase transfection ill conditions where plasmid concentration is low, typically in vivo where mobility of DNA is limited by the extracellular matrix. LV pulses provide additional electrophoretic force which drags DNA toward the cell membrane and consequently increase transfection efficiency, while for sufficiently high concentrations of the plasmid (Usually used in vitro) electrophoretic LV pulses do not have an important role. (C) 2008 Elsevier B.V. All rights reserved.