An Efficient Workflow for CHO Cell Genome Engineering with OpenCRISPR-1

Kolakada, D.; Ankala, R.; Donatelli, J.; Smith, D. K.

doi:10.64898/2025.12.18.695036

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bioRxiv · 10.64898/2025.12.18.695036

An Efficient Workflow for CHO Cell Genome Engineering with OpenCRISPR-1

Kolakada, D.·Ankala, R.·Donatelli, J.·Smith, D. K.

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Abstract

The effective titer and quality of biopharmaceutical products can be enhanced by genome engineering of producer cell lines; however, licensing constraints often limit nuclease utility. Here, we validate OpenCRISPR-1 in CHO cells by achieving [≥]70% INDEL efficiency across multiple genes. We demonstrate biallelic Fut8 knockout in monoclonal cell lines with 31% efficiency and quadruplex knockout of lipases at 7% efficiency using a 39-day workflow. This work highlights the potential applications for democratized nucleases in host cell engineering.

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Kolakada, D., Ankala, R., Donatelli, J., Smith, D. K.. 2025-12-20. An Efficient Workflow for CHO Cell Genome Engineering with OpenCRISPR-1. https://doi.org/10.64898/2025.12.18.695036

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