CRISPR Technologies in Chinese Hamster Ovary Cell Line Engineering

Abstract

The Chinese hamster ovary (CHO) cell line is a well-established platform for the production of biopharmaceuticals due to its ability to express complex therapeutic proteins with human-like glycopatterns in high amounts. The advent of CRISPR technology has opened up new avenues for the engineering of CHO cell lines for improved protein production and enhanced product quality. This review summarizes recent advances in the application of CRISPR technology for CHO cell line engineering with a particular focus on glycosylation modulation, productivity enhancement, tackling adventitious agents, elimination of problematic host cell proteins, development of antibiotic-free selection systems, site-specific transgene integration, and CRISPR-mediated gene activation and repression. The review highlights the potential of CRISPR technology in CHO cell line genome editing and epigenetic engineering for the more efficient and cost-effective development of biopharmaceuticals while ensuring the safety and quality of the final product.

Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

“…CRISPR-Cas9 gene editing technology has been used in CHO cell line engineering to alter the expression of many endogenous genes and produce modified cell lines with the desired phenotype (Kheirandish et al 2023 ; Rahimi et al 2023 ), to improve productivity, growth characteristics and glycosylation, as well as enhance the long-term stability of transgenic expression, and eliminate problematic host cell proteins (HCPs) (Glinsek et al 2023 ).…”

“…Lipoprotein lipase (LPl) has been found to be one of the most common difficult-to-remove HCPs. LPl degrades polysorbate, an excipient commonly used in final drug formulations, negatively affecting the stability of RTPs (Glinsek et al 2023 ). Therefore, LPl is an ideal candidate for gene knockout.…”