A fast and efficient method for transiently transfecting ES cells: application to the development of systems for conditional gene expression.
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.
Bugeon
L
L
Department of Biology, Imperial College of Science Technology and Medicine, London, UK. l.bugeon@ic.ac.uk
Syed
N
N
Dallman
M J
MJ
D002478:Cells, Cultured; D015870:Gene Expression / Q000235:genetics*; D005820:Genetic Testing / Q000379:methods; D007124:Immunoenzyme Techniques; D019426:Integrases / Q000378:metabolism; D007763:Lac Operon; D008081:Liposomes; D012680:Sensitivity and Specificity; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014764:Viral Proteins
D008081:Liposomes; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases