Rapid genetic modification of mouse embryonic stem cells by inducible cassette exchange recombination

Michelina Iacovino, Megan E. Roth, Michael Kyba

Research output: Chapter in Book/Report/Conference proceedingChapter

14 Scopus citations

Abstract

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.

Original languageEnglish (US)
Title of host publicationGene Function Analysis
PublisherHumana Press Inc.
Pages339-351
Number of pages13
ISBN (Print)9781627037204
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1101
ISSN (Print)1064-3745

Keywords

  • Cre
  • ESC
  • ESC targeting
  • Inducible system
  • Recombination

Fingerprint

Dive into the research topics of 'Rapid genetic modification of mouse embryonic stem cells by inducible cassette exchange recombination'. Together they form a unique fingerprint.

Cite this