Engineering RNA export for measurement and manipulation of living cells - Characterizing clonal barcode library diversity

A system for programmable export of RNA molecules from living cells would enable both non-destructive monitoring of cell dynamics and engineering of cells capable of delivering executable RNA programs to other cells. We developed genetically encoded cellular RNA exporters, inspired by viruses, that efficiently and selectively package and secrete target RNA molecules from mammalian cells within protective nanoparticles. Exporting and sequencing RNA barcodes enabled non-destructive monitoring of cell population dynamics with clonal resolution. By incorporating fusogens into the nanoparticles, we demonstrated delivery, expression, and functional activity of mRNA in recipient cells. These engineered RNA export systems thus enable measurement of cell dynamics and establish a foundation for hybrid cell and gene therapies based on cell-to-cell delivery of RNA. This BioProject contains data related to characterization of viral genetic barcode library diversity by DNA sequencing. More specifically, lentiviral libraries containing a high-diversity sequence barcode were generated by molecular cloning, then the barcode region was amplified and sequenced.