Literature DB >> 11032372

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

L Bugeon1, N Syed, M J Dallman.   

Abstract

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.

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Year:  2000        PMID: 11032372     DOI: 10.1023/a:1008990510849

Source DB:  PubMed          Journal:  Transgenic Res        ISSN: 0962-8819            Impact factor:   2.788


  4 in total

1.  Tight control of gene expression in mammalian cells by tetracycline-responsive promoters.

Authors:  M Gossen; H Bujard
Journal:  Proc Natl Acad Sci U S A       Date:  1992-06-15       Impact factor: 11.205

2.  Efficient selection for high-expression transfectants with a novel eukaryotic vector.

Authors:  H Niwa; K Yamamura; J Miyazaki
Journal:  Gene       Date:  1991-12-15       Impact factor: 3.688

3.  Ecdysone-inducible gene expression in mammalian cells and transgenic mice.

Authors:  D No; T P Yao; R M Evans
Journal:  Proc Natl Acad Sci U S A       Date:  1996-04-16       Impact factor: 11.205

4.  Non-injection methods for the production of embryonic stem cell-embryo chimaeras.

Authors:  S A Wood; N D Allen; J Rossant; A Auerbach; A Nagy
Journal:  Nature       Date:  1993-09-02       Impact factor: 49.962

  4 in total
  3 in total

1.  Functional redundancy of GSK-3alpha and GSK-3beta in Wnt/beta-catenin signaling shown by using an allelic series of embryonic stem cell lines.

Authors:  Bradley W Doble; Satish Patel; Geoffrey A Wood; Lisa K Kockeritz; James R Woodgett
Journal:  Dev Cell       Date:  2007-06       Impact factor: 12.270

2.  Delivery of the Cre recombinase by a self-deleting lentiviral vector: efficient gene targeting in vivo.

Authors:  A Pfeifer; E P Brandon; N Kootstra; F H Gage; I M Verma
Journal:  Proc Natl Acad Sci U S A       Date:  2001-09-11       Impact factor: 11.205

3.  A simple and efficient method for transfecting mouse embryonic stem cells using polyethylenimine.

Authors:  Colleen M Bartman; Jennifer Egelston; Xiaojun Ren; Raibatak Das; Christopher J Phiel
Journal:  Exp Cell Res       Date:  2014-08-04       Impact factor: 3.905

  3 in total

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