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

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Article in Methods in molecular biology (Clifton, N.J.), 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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5 · Who and what money

Authors and funding

5 authors.

Lise Marie GravDepartment of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, Denmark. lgrav@dtu.dk.

Johan Blatt RojekDepartment of Biotechnology and Biomedicine, Technical University of Denmark, Lyngby, Denmark.

Karen Julie la Cour KarottkiDepartment of Pediatrics, University of California, San Diego, La Jolla, CA, USA.

Jae Seong LeeDepartment of Molecular Science and Technology, Ajou University, Suwon, Republic of Korea.

Helene Faustrup KildegaardThe Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Lyngby, Denmark.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

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.

Indexed as

CricetulusCRISPR-Cas SystemsGene EditingGlutamate-Ammonia LigaseRecombinant ProteinsAnimalsCHO CellsGene Knockout TechniquesRNA, Guide, CRISPR-Cas SystemsGlutamate-Ammonia LigaseRecombinant ProteinsRNA, Guide, CRISPR-Cas SystemsCas9Chinese hamster ovary (CHO) cellsCRISPRGenome editingGlutamine synthetaseKnockoutRecombinant protein production

Identifiers

PMID39460914

DOI10.1093/nar/gkaa975

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.