Effective gene delivery to adult neurons by a modified form of electroporation

被引：57

作者：

Leclere, PG ^{[1
]}

Panjwani, A ^{[1
]}

Docherty, R ^{[1
]}

Berry, M ^{[1
]}

Pizzey, J ^{[1
]}

Tonge, DA ^{[1
]}

机构：

[1] Kings Coll London, GKT Sch Biomed Sci, Neurosci Res Ctr, London SE1 1UL, England

来源：

JOURNAL OF NEUROSCIENCE METHODS | 2005年 / 142卷 / 01期

基金：

英国生物技术与生命科学研究理事会;

关键词：

retinal ganglion neurons; DRG; transfection; nucleofection; gene transfer; GFP; HSV-1;

D O I：

10.1016/j.jneumeth.2004.08.012

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：137 / 143

页数：7

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