A fast and efficient method for transiently transfecting ES cells: application to the development of systems for conditional gene expression.
作者： Laurence Bugeon , Nelofer Syed , Margaret J. Dallman
关键词:
摘要: Classically, mouse embryonic stem (ES) cells are transfected by electroporation, a method that requires large number of cells. Here we describe protocol using liposome based transfection agent is very simple, rapid and cost effective way transiently transfecting low numbers ES We found this useful in screening clones when working with inducible expression systems which at least two elements required for regulated the gene interest. After stable first component, can be easily rapidly screened interest second component system protocol.
springer.com
cabdirect.org
sci-hub.se 参考文章(4)
Stephen A. Wood, Nick D. Allen, Janet Rossant, Anna Auerbach, Andras Nagy, Non-injection methods for the production of embryonic stem cell-embryo chimaeras Nature. ,vol. 365, pp. 87- 89 ,(1993) , 10.1038/365087A0
D. No, T. P. Yao, R. M. Evans, Ecdysone-inducible gene expression in mammalian cells and transgenic mice Proceedings of the National Academy of Sciences of the United States of America. ,vol. 93, pp. 3346- 3351 ,(1996) , 10.1073/PNAS.93.8.3346
Niwa Hitoshi, Yamamura Ken-ichi, Miyazaki Jun-ichi, Efficient selection for high-expression transfectants with a novel eukaryotic vector Gene. ,vol. 108, pp. 193- 199 ,(1991) , 10.1016/0378-1119(91)90434-D
M. Gossen, H. Bujard, Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proceedings of the National Academy of Sciences of the United States of America. ,vol. 89, pp. 5547- 5551 ,(1992) , 10.1073/PNAS.89.12.5547