A fast and efficient method for transiently transfecting ES cells: application to the development of systems for conditional gene expression

被引：8

作者：

Bugeon, L ^{[1
]}

Syed, N ^{[1
]}

Dallman, MJ ^{[1
]}

机构：

[1] Univ London Imperial Coll Sci Technol & Med, Dept Biol, London SW7 2AZ, England

来源：

TRANSGENIC RESEARCH | 2000年 / 9卷 / 03期

基金：

英国惠康基金;

关键词：

conditional gene expression; ecdysone; embryonic stem cells; gene targeting; liposome; transfection; transgenic mouse;

D O I：

10.1023/A:1008990510849

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Classically, mouse embryonic stem (ES) cells are transfected by electroporation, a method that requires a large number of cells. Here we describe a protocol using a liposome based transfection agent that is a very simple, rapid and cost effective way of transiently transfecting very low numbers of ES cells. We found this method very useful in screening a large number of ES clones when working with inducible expression systems in which at least two elements are required for regulated expression of the gene of interest. After stable transfection of the first component, clones can be easily and rapidly screened for expression of the gene of interest by transiently transfecting the second component of the system using this protocol.

引用

页码：229 / 232

页数：4

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