Efficient transfection of DNA or shRNA vectors into neurons using magnetofection

被引：104

作者：

Buerli, Thomas ^{[2
,3
]}

Pellegrino, Christophe ^{[1
]}

Baer, Kristin ^{[4
]}

Lardi-Studler, Barbara ^{[3
]}

Chudotvorova, Ilona ^{[1
]}

Fritschy, Jean-Marc ^{[3
]}

Medina, Igor ^{[1
]}

Fuhrer, Christian ^{[2
]}

机构：

[1] INSERM, INMED, Unite 29, F-13009 Marseille, France

[2] Univ Zurich, Dept Neurochem, Inst Brain Res, CH-8057 Zurich, Switzerland

[3] Univ Zurich, Inst Pharmacol & Toxicol, CH-8057 Zurich, Switzerland

[4] Univ Coll Swansea, Sch Med, Inst Life Sci, Swansea SA2 8PP, W Glam, Wales

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 12期

关键词：

D O I：

10.1038/nprot.2007.445

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Efficient and long-lasting transfection of primary neurons is an essential tool for addressing many questions in current neuroscience using functional gene analysis. Neurons are sensitive to cytotoxicity and difficult to transfect with most methods. We provide a protocol for transfection of cDNA and RNA interference (short hairpin RNA (shRNA)) vectors, using magnetofection, into rat hippocampal neurons (embryonic day 18/19) cultured for several hours to 21 d in vitro. This protocol even allows double-transfection of DNA into a small subpopulation of hippocampal neurons (GABAergic interneurons), as well as achieving long-lasting expression of DNA and shRNA constructs without interfering with neuronal differentiation. This protocol, which uses inexpensive equipment and reagents, takes 1 h; utilizes mixed hippocampal cultures, a transfection reagent, CombiMag, and a magnetic plate; shows low toxicity and is suited for single-cell analysis. Modifications done by our three laboratories are detailed.

引用

页码：3090 / 3101

页数：12

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