Reverse transfection on cell arrays for high content screening microscopy

被引：155

作者：

Erfle, Holger ^{[1
]}

Neumann, Beate ^{[1
]}

Liebel, Urban ^{[1
]}

Rogers, Phill ^{[1
]}

Held, Michael ^{[1
]}

Walter, Thomas ^{[1
]}

Ellenberg, Jan

Pepperkok, Rainer ^{[2
]}

机构：

[1] European Mol Biol Lab, MitoCheck Project Grp, D-69117 Heidelberg, Germany

[2] European Mol Biol Lab, Cell Biol Biophys Programmes, D-69117 Heidelberg, Germany

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 02期

关键词：

D O I：

10.1038/nprot.2006.483

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Here, we describe a robust protocol for the reverse transfection of cells on small interfering (siRNA) arrays, which, in combination with multi-channel immunofluorescence or time-lapse microscopy, is suitable for genome-wide RNA interference (RNAi) screens in intact human cells. The automatic production of 48 'transfection ready' siRNA arrays, each containing 384 samples, takes in total 7 h. Pre-fabricated siRNA arrays can be used without loss of transfection efficiency at least up to 15 months after printing. Different human cell lines that have been successfully transfected using the protocol are presented here. The present protocol has been applied to two genome-wide siRNA screens addressing mitosis and constitutive protein secretion.

引用

页码：392 / 399

页数：8

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