Rapid genetic modification of mouse embryonic stem cells by Inducible Cassette Exchange recombination.
Embryonic stem cell (ESC) differentiation is a useful means by which to produce large quantities of cells in vitro representing early stages of embryonic development. A conditional gene expression system allows interrogation of factors at specific time points in the differentiation of ES cells to defined cell types. We have developed a method for rapidly generating conditional inducible murine ES cells by targeting genes into an Inducible Cassette Exchange (ICE) locus. The ICE locus encodes a doxycycline-inducible floxed Cre, which replaces itself with an incoming floxed gene of interest. The derivative cell lines, selected in G418, thus bear doxycycline-inducible transgenes. We provide detailed methods for performing ICE recombination and generating derivative doxycycline-inducible cell lines.
Department of Pediatrics, Harbor-UCLA Medical Center, Torrance, CA, USA.
Michelina
M
Iacovino
Megan E
ME
Roth
Michael
M
Kyba
D000818:Animals; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000502:physiology*; D061252:Feeder Cells; D060005:Genotyping Techniques; D019426:Integrases / Q000235:genetics; D051379:Mice; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D015533:Transcriptional Activation; D019076:Transgenes
C045073:Cre recombinase; D019426:Integrases