biostudies-literatureUnknown12Zhang CSergey Brin Family FoundationHoward Hughes Medical InstituteInnovative Genomics Institute, UC BerkeleyMuch attention has been focused on the possibility that cytoplasmic proteins and RNA may be conveyed between cells in extracellular vesicles (EVs) and tunneling nanotube (TNT) structures. Here, we set up two quantitative delivery reporters to study cargo transfer between cells. We found that EVs are internalized by reporter cells but do not efficiently deliver functional Cas9 protein to the nucleus. In contrast, donor and acceptor cells co-cultured to permit cell contact resulted in a highly effective transfer. Among our tested donor and acceptor cell pairs, HEK293T and MDA-MB-231 recorded optimal intercellular transfer. Depolymerization of F-actin greatly decreased Cas9 transfer, whereas inhibitors of endocytosis or knockdown of genes implicated in this process had little effect on transfeLifee84391https://www.ebi.ac.uk/biostudies/studies/S-EPMC10112890biostudies-literatureSyncytin-mediated open-ended membrane tubular connections facilitate the intercellular transfer of cargos including Cas9 protein.PMC10112890Schekman RZhang CfalseSyncytin-mediated open-ended membrane tubular connections facilitate the intercellular transfer of cargos including Cas9 protein.Much attention has been focused on the possibility that cytoplasmic proteins and RNA may be conveyed between cells in extracellular vesicles (EVs) and tunneling nanotube (TNT) structures. Here, we set up two quantitative delivery reporters to study cargo transfer between cells. We found that EVs are internalized by reporter cells but do not efficiently deliver functional Cas9 protein to the nucleus. In contrast, donor and acceptor cells co-cultured to permit cell contact resulted in a highly effective transfer. Among our tested donor and acceptor cell pairs, HEK293T and MDA-MB-231 recorded optimal intercellular transfer. Depolymerization of F-actin greatly decreased Cas9 transfer, whereas inhibitors of endocytosis or knockdown of genes implicated in this process had little effect on transf2023-01-01T00:00:00Z2023 Mar2026-05-28T17:59:09.289Z2024-12-03T15:59:59.564ZS-EPMC101128903689679110.7554/eLife.84391