Cellular microvesicle pathways can be targeted to transfer genetic information between non-immune cells.
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 degrees 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 degrees target cells.
Skinner
Amy M
AM
Papé Family Pediatric Research Institute, Oregon Health & Science University, Portland, OR, USA.
O'Neill
S Lee
SL
Kurre
Peter
P
D001483:Base Sequence; D015153:Blotting, Western; D002450:Cell Communication; D002460:Cell Line; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers