FUT8knockout

FUT8knockout refers to the genetic disruption or inactivation of the FUT8 gene, which encodes the alpha1,6-fucosyltransferase responsible for core fucosylation of N-glycans in vertebrates. In mammals, most N-linked glycoproteins acquire a core fucose on the innermost N-acetylglucosamine of the glycan core, a modification driven by FUT8.

FUT8 encodes the enzyme that adds a fucose residue in an alpha1,6 linkage to N-glycan cores during glycoprotein maturation in the secretory pathway. Core fucosylation influences the structure and recognition properties of glycoproteins, affecting interactions with lectins, receptors, and antibodies. Because of its central role, FUT8 activity modulates processes such as cell signaling, adhesion, and immune recognition.

In FUT8 knockout systems, core fucosylation is abolished or greatly reduced. This leads to altered glycan profiles on many glycoproteins, with notable functional consequences. A well-documented outcome is enhanced antibody-dependent cellular cytotoxicity (ADCC) mediated by afucosylated antibodies, due to improved binding to Fc gamma receptors. Knockout models also provide insight into the role of core fucosylation in development and physiology, though effects can vary between species and tissues.

Applications include biotechnology and basic research. FUT8 knockout cell lines are used to produce afucosylated therapeutic antibodies with higher effector activity. Researchers use FUT8-deficient models to study the contributions of core fucosylation to protein function, receptor signaling, and disease mechanisms.

Methods to create FUT8 knockouts typically involve CRISPR/Cas9, TALENs, or homologous recombination, with validation by mass spectrometry of N-glycans, lectin binding assays, and sequencing.