Improving recombinant protein production in CHO cells using the CRISPR-Cas system

Ali Kerem Kalkan, Fahreddin Palaz, Semeniuk Sofija, Nada Elmousa, Yuri Ledezma, Elise Cachat, Leonardo Rios-Solis

Research output: Contribution to journalReview articlepeer-review

Abstract / Description of output

Chinese hamster ovary (CHO) cells are among the most widely used mammalian cell lines in the biopharmaceutical industry. Therefore, it is not surprising that significant efforts have been made around the engineering of CHO cells using genetic engineering methods such as the CRISPR-Cas system. In this review, we summarize key recent studies that have used different CRISPR-Cas systems such as Cas9, Cas13 or dCas9 fused with effector domains to improve recombinant protein (r-protein) production in CHO cells. Here, every relevant stage of production was considered, underscoring the advantages and limitations of these systems, as well as discussing their bottlenecks and probable solutions. A special emphasis was given on how these systems could disrupt and/or regulate genes related to glycan composition, which has relevant effects over r-protein properties and in vivo activity. Furthermore, the related promising future applications of CRISPR to achieve a tunable, reversible, or highly stable editing of CHO cells are discussed.

Overall, the studies covered in this review show that despite the complexity of mammalian cells, the synthetic biology community has developed many mature strategies to improve r-protein production using CHO cells. In this regard, CRISPR-Cas technology clearly provides efficient and flexible genetic manipulation and allows for the generation of more productive CHO cell lines, leading to more cost-efficient production of biopharmaceuticals, however, there is still a need for many emerging techniques in CRISPR to be reported in CHO cells; therefore, more research in these cells is needed to realize the full potential of this technology.
Original languageEnglish
Article number108115
Number of pages18
JournalBiotechnology Advances
Publication statusPublished - 7 Feb 2023

Keywords / Materials (for Non-textual outputs)

  • CRISPR system
  • CHO cells
  • recombinant protein
  • bioprocessing
  • glycan composition
  • synthetic Biology
  • mammalian cells


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