CRISPR-Cas Systems and CHO in Biopharmaceuticals: Unlocking NewPossibilities in Gene Editing

The CRISPR-Cas system has significantly advanced genome editing, offering superior efficiency, precision, and ease of use compared to traditional technologies such as Zinc Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs). CHO cells are a widely used mammalian cell line for large-scale therapeutic protein manufacturing due to their ability to produce human-like glycosylation patterns and grow in serum-free media. Recent CRISPR-based CHO cell engineering enables precise genetic modifications, improving productivity, stability, scalability, and reducing costs. This article highlights the transformative role of CRISPR technologies in addressing genetic disorders and expanding the frontiers of multiple scientific fields. It offers a comprehensive analysis of several CRISPR-Cas systems, including Cas9, Cas12, Cas13, and Cas14, emphasizing their unique structural features and functional capabilities. While Cas9 has dominated many genomeediting applications, the use of Cas13 in Chinese Hamster Ovary (CHO) cells has opened up promising RNA-targeting strategies. Moreover, the compact Cas14 system presents notable potential for applications requiring ultra-precise genome manipulation. With their critical role in therapeutic protein production, CHO cells have greatly benefited from CRISPR-enabled engineering, leading to measurable improvements in productivity, stability, and cost-efficiency. Key advancements in CRISPR delivery platforms, including both viral and nonviral vectors, are discussed alongside ongoing challenges such as off-target effects and regulatory considerations. Emerging trends such as base editing, prime editing, and the integration of artificial intelligence for system optimization are also explored. Altogether, the discussion underscores the pivotal contribution of CRISPR technologies to CHO cell engineering and their broader impact on the future of biopharmaceutical manufacturing