Chinese hamster ovary (CHO) cells are the preferred workhorse for the biopharmaceutical industry, and CRISPR/Cas9 has proven powerful for generating targeted gene perturbations in CHO cells. Here, we expand the CRISPR engineering toolbox with CRISPR activation (CRISPRa) to increase transcription of endogenous genes. We successfully increased transcription of Mgat3 and St6gal1, and verified their activity on a functional level by subsequently detecting that the appropriate glycan structures were produced. This study demonstrates that CRISPRa can make targeted alterations of CHO cells for desired phenotypes.
The authors thank Helle Munck Petersen and Sanne Schoffelen for help with glycosylation labeling. The authors also acknowledge support from the Novo Nordisk Foundation (NNF10CC1016517 and NNF16OC0021638) and NIGMS (R35 GM119850).
