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Targeted Mutations in the Mouse via Embryonic Stem Cells

  • Marina GertsensteinEmail author
  • Joffrey Mianné
  • Lydia Teboul
  • Lauryl M. J. Nutter
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2066)

Abstract

Genetic modification of mouse embryonic stem (ES) cells is a powerful technology that enabled the generation of a plethora of mutant mouse lines to study gene function and mammalian biology. Here we describe ES cell culture and transfection techniques used to manipulate the ES cell genome to obtain targeted ES cell clones. We include the standard gene targeting approach as well as the application of the CRISPR/Cas9 system that can improve the efficiency of homologous recombination in ES cells by introducing a double-strand DNA break at the target site.

Key words

Mouse Embryonic stem (ES) cells Gene targeting Electroporation Clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR associated protein (Cas9) 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Marina Gertsenstein
    • 1
    Email author
  • Joffrey Mianné
    • 2
  • Lydia Teboul
    • 2
  • Lauryl M. J. Nutter
    • 1
    • 3
  1. 1.The Centre for Phenogenomics (TCP)TorontoCanada
  2. 2.The Mary Lyon CentreMRC Harwell InstituteDidcotUK
  3. 3.Genetics and Genome BiologyThe Hospital for Sick ChildrenTorontoCanada

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