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

Lise Marie Grav, Karen Julie la Cour Karottki, Jae Seong Lee, Helene Faustrup Kildegaard

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Abstract

Genome editing has become an increasingly important aspect of Chinese Hamster Ovary (CHO ) cell line engineering for improving production of recombinant protein therapeutics. Currently, the focus is directed toward expanding the product diversity, controlling and improving product quality and yields. In this chapter, we present our protocol on how to use the genome editing tool Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) to knockout engineering target genes in CHO cells. As an example, we refer to the glutamine synthetase (GS)-encoding gene as the knockout target gene, a knockout that increases the selection efficiency of the GS-mediated gene amplification system.
Original languageEnglish
Title of host publicationHeterologous Protein Production in CHO cells
PublisherSpringer
Publication date2017
Pages101-118
Chapter7
DOIs
Publication statusPublished - 2017
SeriesMethods in Molecular Biology
Volume1603
ISSN1064-3745

Keywords

  • CRISPR/Cas9
  • Chinese Hamster Ovary Cells
  • Genome editing
  • Glutamine synthetase
  • Knockout
  • Recombinant protein production

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