Cell-based glycoengineering of extracellular vesicles through precise genome editing

• DOI: 10.1016/j.nbt.2024.07.004
• Publication Date: 2024-07-28T05:34:36Z
• Authors: Weihua Tian; Chiara Zagami; Jiasi Chen; Anne Louise Blomberg; Laura Salse Guiu; Sarah Line Skovbakke; Steffen Goletz
• Keywords: 0301 basic medicine; 03 medical and health sciences; Glycoengineering; Carbohydrates; Small extracellular vesicles; EV glycobiology; Gene editing

Engineering of extracellular vesicles (EVs) towards more efficient targeting and uptake to specific cells has large potentials for their application as therapeutics. Carbohydrates play key roles in various biological interactions are essential EV biology. The extent which glycan modification EVs can be achieved through genetic glycoengineering parental not been explored yet. Here we introduce targeted cell-based via enzymes the glycosylation machinery. In a "simple cell" strategy, modified major pathways by knocking-out (KO) genes N-glycosylation (MGAT1), O-GalNAc (C1GALT1C1), glycosphingolipids (B4GALT5/6), glycosaminoglycans (B4GALT7) sialylation (GNE) involved elongation or biosynthesis glycans HEK293F cells. gene editing led corresponding changes on demonstrated differential lectin staining. Small (sEVs) isolated from showed overall changes, but also some unexpected differences cell including enrichment preference certain structures absence other types. did significantly impact sEVs production, size distribution, syntenin-1 biomarker expression, while clonal influence production yields was observed. Our findings demonstrate successful implementation stable source generation glycoengineered sEVs. utilization offers promising opportunity study role biology, well facilitate optimization therapeutic purposes.