FUT8 Knockout HEK293F Cell Line

The FUT8 Knockout HEK293F Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the suspension-adapted HEK293F host. This model carries a targeted disruption of the FUT8 gene, which encodes ??1,6-fucosyltransferase, the enzyme that catalyzes core fucose addition to N-linked glycans. The stable loss-of-function cell line is particularly suited for glycoengineering studies and the bioproduction of afucosylated antibodies for enhanced ADCC. The knockout is achieved using CRISPR/Cas9-mediated gene disruption, ensuring heritable ablation of core fucosylation without reliance on transient inhibitors or knockdown approaches.

HEK293F cells are a suspension-adapted derivative of the widely used HEK293 line, originally established by adenovirus type 5 transformation of human embryonic kidney tissue. These cells are optimized for high-density culture in serum-free media and are extensively employed for transient and stable production of recombinant proteins and viral vectors. Their human origin ensures proper folding and post-translational modifications, making them a preferred host for therapeutic protein manufacturing. The suspension growth format enables scalable bioreactor operation, ideal for bioprocess development.

FUT8 encodes the sole ??1,6-fucosyltransferase in humans, catalyzing core fucosylation of N-glycans using GDP-fucose. This modification modulates glycoprotein stability, cell adhesion, and antibody effector functions. FUT8 expression is activated by TGF-??1/Smad2/3 and EGF signaling. Downstream, core fucosylation of E-cadherin and integrins impacts cell?Cmatrix interactions, while fucosylation of the IgG Fc domain sterically inhibits binding to Fc??RIIIa, diminishing antibody-dependent cellular cytotoxicity (ADCC). Genetic disruption of FUT8 removes core fucose, enhancing Fc??RIIIa engagement and ADCC potency.

In the HEK293F background, disruption of FUT8 directly prevents core fucosylation of recombinant glycoproteins, most prominently antibodies. This yields uniformly afucosylated products with markedly improved ADCC activity, a critical attribute for therapeutic antibodies targeting cancer and autoimmune diseases. The suspension growth capability combined with FUT8 knockout enables scalable bioproduction of glycoengineered antibodies with enhanced effector functions. Moreover, absence of core fucose simplifies analytical glycoprofiling and quality control of manufactured glycoproteins, while also facilitating cell adhesion and signaling studies.

This cell line is suited for bioproduction of afucosylated antibodies for functional ADCC assays, investigation of core fucosylation in EGFR, integrin, and E-cadherin signaling, and glycoengineering studies using lectin blotting, flow cytometry with fucose-binding lectins (e.g., AAL or LCA), and LC-MS glycoprofiling. It also serves as a model for congenital disorders of glycosylation and for exploring how FUT8 contributes to cancer metastasis and immune evasion. For further information, please contact Ascent Research.