Mammalian cells lacking either the cotranslational or posttranslocational oligosaccharyltransferase complex display substrate-dependent defects in asparagine linked glycosylation.

摘要: Asparagine linked glycosylation of proteins is an essential protein modification reaction in most eukaryotic organisms. Metazoan organisms express two oligosaccharyltransferase complexes that are composed a catalytic subunit (STT3A or STT3B) assembled with shared set accessory subunits and one to complex specific subunits. siRNA mediated knockdowns STT3A STT3B HeLa cells have shown the OST partially non-overlapping roles N-linked glycosylation. However, incomplete depletion reduces impact loss, thereby complicating interpretation experimental results. Here, we used CRISPR/Cas9 gene editing technology create viable HEK293 derived lines deficient for single STT3B-specific (MagT1 TUSC3). Analysis STT3A, MagT1/TUSC3 null cell revealed these superior tools investigating vivo role substrate preferences complexes.