Bi-allelic gene targeting in mouse embryonic stem cells

Methods. 2011 Apr;53(4):331-8. doi: 10.1016/j.ymeth.2010.12.025. Epub 2011 Feb 1.

Abstract

The EUCOMM and KOMP programs have generated targeted conditional alleles in mouse embryonic stem cells for nearly 10,000 genes. The availability of these stem cell resources will greatly accelerate the functional analysis of genes in mice and in cultured cells. We present a method for conditional ablation of genes in ES cells using vectors and targeted clones from the EUCOMM and KOMP conditional resources. Inducible homozygous cells described here provide a precisely controlled experimental system to study gene function in a model cell.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles*
  • Animals
  • Autoantigens / genetics
  • Cell Culture Techniques
  • Cell Line
  • Cloning, Molecular / methods
  • DNA Nucleotidyltransferases / genetics
  • Electroporation / methods
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / physiology*
  • Gene Expression Regulation
  • Gene Targeting / methods*
  • Genetic Vectors / biosynthesis
  • Genotype
  • Homozygote
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex / genetics
  • Mice
  • Mice, Inbred C57BL
  • Polymerase Chain Reaction / methods
  • Promoter Regions, Genetic
  • Transcription, Genetic
  • Transfection / methods

Substances

  • Autoantigens
  • CHD4 protein, human
  • DNA Nucleotidyltransferases
  • FLP recombinase
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex