Efficient multiplexed targeted gene editing as a strategy to generate improved CHO host cell lines for biotherapeutic manufacturing applications

The use of CRISPR-Cas9 tools for gene editing has opened new possibilities for the genetic optimization of biological systems with biotechnological or therapeutic applications. One area of interest is the design of enhanced expression platforms and the generation of new host cell lines that have the required phenotypic advantages and performance for use in biopharmaceutical manufacturing. However, the long-term success of generating new host cells depends greatly on the ability to perform multiplexed gene editing with high efficiency and the deployment of suitable quality control methodologies to accurately evaluate all editing outcomes.

In this whitepaper, we demonstrate Revvity’s workflow exemplifying the successful simultaneous knock-out of multiple endogenous loci in CHO cells and show the absence of the targeted proteins in several of the selected clones, proving the efficiency of the multiplexed gene editing approach. It’s also shown that simultaneous knock-out of multiple loci using CRISPR-Cas9 and multiple gRNAs is an effective strategy to enhance CHO cell protein production levels.

In collaboration with Cergentis, read more about the data generated including:

• Gene copy number analysis and screening for gene editing efficiency
• Generation and characterization of an edited pool
• Isolation and phenotypic validation of edited clones
• Targeted Locus Amplification

The CHOSOURCE™ Platform is available for research, clinical, diagnostic, and commercialization applications under specific licenses from Revvity. The platform is also available for services under a service license from Revvity.