Abstract
Eukaryotic cell communication is based on protein signaling cascades that require direct cell-cell apposition, or receptor engagement by secreted molecules. The transmission of genetic information is thought to be uncommon, apart from recent reports of exosomal RNA transfer in immune and glioblastoma cells. We wished to examine if existing microvesicle pathways could be directly targeted for the horizontal transfer of RNA genomes in less specialized cell types. Using replication-deficient retrovirus vector, studies herein confirm that a range of cells routinely sequester a small population of these RNA genomes in a non-canonical compartment, refractory to antibody neutralization and unaffected by specific pharmacological inhibition of pathways involved in conventional viral trafficking. Our experiments further reveal the cytoplasmic colocalization of vector genomes with tetraspanin proteins as well as the PI-3-kinase sensitive trafficking and subsequent transmission to 2° targets. Collectively, our results indicate a scalable process whereby cells route vector genomes to multivesicular bodies (MVB) for cytoplasmic trafficking and exosomal release. Our findings imply that cells can serve to deliver recombinant payload, targeted for the stable genetic modification of 2° target cells.
Highlights
Eukaryotic cell communication is based on protein signaling via direct cell-cell contacts, or indirectly via ligand-receptor interactions
Recent work suggests that cell-cell communication may occur in part through transfer in membrane-derived vesicles that stem from the fusion of multivesicular bodies (MVB) with the plasma membrane [1]
We evaluated the intracellular location of GFP expressing vector particles (GFP-vpr) tagged vector genomes and their colocalization with select endosomal compartments: AP2, EEA1, H68.4, and N-Rh-PE
Summary
Eukaryotic cell communication is based on protein signaling via direct cell-cell contacts, or indirectly via ligand-receptor interactions. The recently described microvesicle transfer of RNA between glioblastoma cells or the exosomal cell-cell transmission of microRNA in mast cells provide highly specialized exceptions of ‘‘horizontal’’ genetic communication among target cells [2,3]. Those studies demonstrate microvesicle mediated transfer and cytoplasmic detection of donor cell ‘‘RNA signatures’’ in 2u targets. Little is known about the recruitment and trafficking of RNA to such a pathway and its potential existence in less specialized cell populations. There have been no demonstrations of long-lived effects in 2u targets, nor attempts to directly exploit such genetic communication
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