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Review
. 2023 Apr;120(4):865-902.
doi: 10.1002/bit.28329. Epub 2023 Jan 12.

CRISPR-interceded CHO cell line development approaches

Shahin Amiri  1 , Setare Adibzadeh  1 , Samaneh Ghanbari  1 , Behnaz Rahmani  1 , Mohammad H Kheirandish  1   2 , Aref Farokhi-Fard  1 , Mansoureh S Dastjerdeh  1 , Fatemeh Davami  1
Affiliations
Review

CRISPR-interceded CHO cell line development approaches

Shahin Amiri et al. Biotechnol Bioeng. 2023 Apr.

Abstract

For industrial production of recombinant protein biopharmaceuticals, Chinese hamster ovary (CHO) cells represent the most widely adopted host cell system, owing to their capacity to produce high-quality biologics with human-like posttranslational modifications. As opposed to random integration, targeted genome editing in genomic safe harbor sites has offered CHO cell line engineering a new perspective, ensuring production consistency in long-term culture and high biotherapeutic expression levels. Corresponding the remarkable advancements in knowledge of CRISPR-Cas systems, the use of CRISPR-Cas technology along with the donor design strategies has been pushed into increasing novel scenarios in cell line engineering, allowing scientists to modify mammalian genomes such as CHO cell line quickly, readily, and efficiently. Depending on the strategies and production requirements, the gene of interest can also be incorporated at single or multiple loci. This review will give a gist of all the most fundamental recent advancements in CHO cell line development, such as different cell line engineering approaches along with donor design strategies for targeted integration of the desired construct into genomic hot spots, which could ultimately lead to the fast-track product development process with consistent, improved product yield and quality.

Keywords: CRISPR-Cas9; Chinese hamster ovary; cell line development; donor design; targeted integration.

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References

REFERENCES

    1. Ahmadi, M., Mahboudi, F., Akbari Eidgahi, M. R., Nasr, R., Nematpour, F., Ahmadi, S., Ebadat, S., Aghaeepoor, M., & Davami, F. (2016). Evaluating the efficiency of phiC31 integrase-mediated monoclonal antibody expression in CHO cells. Biotechnology Progress, 32(6), 1570-1576. https://doi.org/10.1002/btpr.2362
    1. Aird, E. J., Lovendahl, K. N., St. Martin, A., Harris, R. S., & Gordon, W. R. (2018). Increasing Cas9-mediated homology-directed repair efficiency through covalent tethering of DNA repair template. Communications Biology, 1(1), 54. https://doi.org/10.1038/s42003-018-0054-2
    1. Aldrich, T. L., Viaje, A., & Morris, A. E. (2003). EASE vectors for rapid stable expression of recombinant antibodies. Biotechnology Progress, 19(5), 1433-1438. https://doi.org/10.1021/bp034056j
    1. Amann, T., Hansen, A. H., Kol, S., Hansen, H. G., Arnsdorf, J., Nallapareddy, S., Voldborg, B., Lee, G. M., Andersen, M. R., & Kildegaard, H. F. (2019). Glyco-engineered CHO cell lines producing alpha-1-antitrypsin and C1 esterase inhibitor with fully humanized N-glycosylation profiles. Metabolic Engineering, 52, 143-152. https://doi.org/10.1016/j.ymben.2018.11.014
    1. Amann, T., Hansen, A. H., Kol, S., Lee, G. M., Andersen, M. R., & Kildegaard, H. F. (2018). CRISPR/Cas9-multiplexed editing of Chinese hamster ovary B4Gal-T1, 2, 3, and 4 tailors N-glycan profiles of therapeutics and secreted host cell proteins. Biotechnology Journal, 13(10), 1800111. https://doi.org/10.1002/biot.201800111

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