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Measuring Recombination Proficiency in Mouse Embryonic Stem Cells

  • Andrew J. Pierce
  • Maria Jasin
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1105)

Abstract

A method is presented to measure homologous recombination in mouse embryonic stem cells by both gene targeting and short-tract gene conversion of a double-strand break (DSB). A fluorescence-based reporter is first gene targeted to the Hprt locus in a quantifiable way. A homing endonuclease expression vector is then introduced to generate a DSB, the repair of which is also quantifiable.

Key words

Recombination Double-strand break (DSB) Hprt Mouse embryonic stem (ES) cells Green fluorescent protein (GFP) Flow cytometry Gene targeting Gene conversion I-SceHoming endonuclease 

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Copyright information

© Humana Press 2014

Authors and Affiliations

  1. 1.Cell Biology ProgramMemorial Sloan-Kettering Cancer Center, Cornell University Graduate School of Medical SciencesNew YorkUSA
  2. 2.Markey Cancer CenterUniversity of KentuckyLexingtonUSA

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