CRISPR/Cas9 as a Genome Editing Tool for Targeted Gene Integration in CHO Cells

  • Daria Sergeeva
  • Jose Manuel Camacho-Zaragoza
  • Jae Seong Lee
  • Helene Faustrup KildegaardEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1961)


The emergence of CRISPR/Cas9 system as a precise and affordable method for genome editing has prompted its rapid adoption for the targeted integration of transgenes in Chinese hamster ovary (CHO) cells. Targeted gene integration allows the generation of stable cell lines with a controlled and predictable behavior, which is an important feature for the rational design of cell factories aimed at the large-scale production of recombinant proteins. Here we present the protocol for CRISPR/Cas9-mediated integration of a gene expression cassette into a specific genomic locus in CHO cells using homology-directed DNA repair.

Key words

Chinese hamster ovary cells CRISPR/Cas9 Genome editing Targeted integration 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Daria Sergeeva
    • 1
  • Jose Manuel Camacho-Zaragoza
    • 1
  • Jae Seong Lee
    • 2
  • Helene Faustrup Kildegaard
    • 1
    Email author
  1. 1.The Novo Nordisk Foundation Center for BiosustainabilityTechnical University of DenmarkKgs. LyngbyDenmark
  2. 2.Department of Molecular Science and TechnologyAjou UniversitySuwonRepublic of Korea

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