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

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.