Effective gene delivery to adult neurons by a modified form of electroporation
Effective gene delivery to adult neurons by a modified form of electroporation
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DOI：
10.1016/j.jneumeth.2004.08.012
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发表时间：
2005-03-15
影响因子：
3
通讯作者：
Tonge, DA
中科院分区：
文献类型：
--
作者：
Leclere, PG;Panjwani, A;Tonge, DA
将cDNA转染到成年神经元和其他有丝分裂后细胞中的非病毒方法通常效率很低。然而，由Amaxa Biosystems开发的Nucleofector（TM）技术的最新发展允许将cDNA直接递送到细胞核中，使得能够转染非分裂细胞。在这项研究中，我们描述了一种可靠的方法，培养大量的视网膜细胞从成年大鼠和使用核转染，我们能够转染的cDNA编码的绿色荧光蛋白（水母绿色荧光蛋白）到视网膜神经节细胞（RGCs）与相对较高的效率（高达28%）。在18小时内观察到神经元GFP表达，并持续长达14天。这与用HSV-1载体感染后表达GFP的RGC高达60%的值相比。成年大鼠背根神经节（DRG）神经元也成功转染。总之，核转染为成年哺乳动物神经元中cDNA的递送和基因功能的研究提供了一种快速有效的方法。(C)2004 Elsevier B. V.保留所有权利。
Non-viral methods of transfection of cDNAs into adult neurons and other post-mitotic cells are generally very inefficient. However, the recent development of Nucleofector(TM) technology developed by Amaxa Biosystems allows direct delivery of cDNAs into the nucleus, enabling transfection of non-dividing cells. In this study, we describe a reliable method for culturing large numbers of retinal cells from adult rats and using Nucleofection, we were able to transfect cDNA-encoding GFP (jellyfish green fluorescent protein) into retinal ganglion cells (RGCs) with relatively high efficiency (up to 28%). Neuronal GFP expression was observed within 18 h and continued for up to 14 days. This compares with values up to 60% of RGCs expressing GFP following infection with an HSV-1 vector. Adult rat dorsal root ganglion (DRG) neurons were also successfully transfected. Thus, in summary, Nucleofection provides the possibility for a fast and efficient method for cDNA delivery and study of gene function in adult mammalian neurons. (C) 2004 Elsevier B.V. All rights reserved.