Application of CRISPR/Cas9 Genome Editing to Improve Recombinant Protein Production in CHO Cells

Lise Marie Grav*, Johan Blatt Rojek, Karen Julie la Cour Karottki, Jae Seong Lee, Helene Faustrup Kildegaard

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingBook chapterEducationpeer-review

Abstract

Genome editing has become an important aspect of Chinese hamster ovary (CHO) cell line engineering for improving the production of recombinant protein therapeutics. Currently, the engineering focus is directed toward expanding product diversity while controlling and improving product quality and yields. In this chapter, we present our protocol for using the genome editing tool Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) to knock out engineering target genes in CHO cells. As an example, we describe how to knock out the glutamine synthetase (GS) gene, which increases the selection efficiency of the GS-mediated gene amplification system.
Original languageEnglish
Title of host publicationHeterologous Protein Production in CHO Cells : Methods and Protocols
EditorsPaula Meleady
PublisherSpringer
Publication date2025
Pages49-69
Chapter5
ISBN (Print)978-1-0716-4106-4, 978-1-0716-4103-3
ISBN (Electronic)978-1-0716-4104-0
DOIs
Publication statusPublished - 2025
SeriesMethods in Molecular Biology
Number2853
ISSN1064-3745

Keywords

  • Chinese hamster ovary (CHO) cells
  • CRISPR
  • CAS9
  • Genome editing
  • Glutamine synthetase
  • Knockout
  • Recombiant protein protection

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