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

Kalkan, Ali Kerem; Palaz, Fahreddin; Sofija, Semeniuk; Elmousa, Nada; Ledezma, Yuri; Cachat, Elise; Rios-Solis, Leonardo; (2023) Improving recombinant protein production in CHO cells using the CRISPR-Cas system. Biotechnology Advances , 64 , Article 108115. 10.1016/j.biotechadv.2023.108115. Green open access

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Abstract

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.

Type: Article
Title: Improving recombinant protein production in CHO cells using the CRISPR-Cas system
Location: England
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.biotechadv.2023.108115
Publisher version: https://doi.org/10.1016/j.biotechadv.2023.108115
Language: English
Additional information: Copyright © 2023 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Keywords: CRISPR system; CHO cells; Recombinant protein; Bioprocessing; Glycan composition; Synthetic biology; Mammalian cell
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10183080
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