Fig 3. Flanking mRNA with MmPeg10 5’ and 3’ UTRs enables functional intercellular transfer of mRNA into a target cell.

A. Schematic showing reprogramming MmPEG10 for functional delivery of a cargo RNA flanked with the MmPeg10 5′ and 3′ UTRs (hereafter, “cargo(RNA)”)

B epresentative TEM micrographs of VLP fraction immunogold labeled for MmPEG10. Text labels indicate transfection of cells with MmPeg10 or mock (negative). Arrowheads indicate gold labeling. Scale bar, 50 nm.

C. Representative images of loxP-GFP N2a cells treated with VSVg pseudotyped MmPEG10 VLPs produced by transfecting Mm.cargo(Cre) or Cre mRNA, and a lentivirus encoding Cre. Scale bar, 100 um.

D. Functional transfer of RNA into loxP-GFP N2a cells mediated by VSVg pseudotyped MmPEG10 VLPs. Data quantified by flow cytometry 72 hours after VLP addition, n = 3 replicates.

E. Functional transfer of RNA into loxP-GFP N2a cells mediated by VSVg pseudotyped VLPs produced with MmPeg10 or mCherry and Mm.cargo(Cre) constructs encoding tiles of the MmPeg10 3’UTR. Data quantified by flow cytometry 72 hours after VLP addition, n = 3 replicates.

F. Functional transfer of RNA into loxP-GFP N2a cells mediated by VSVg pseudotyped VLPs produced with HsPEG1010 or mCherry and Hs.cargo(Cre) constructs encoding tiles of the HsPeg10 3’UTR. Data quantified by flow cytometry 72 hours after VLP addition, n = 3 replicates.

G. Functional transfer of RNA into loxP-GFP N2a cells mediated by VSVg pseudotyped VLPs produced with rMmPeg10 and Mm.cargo(Cre) or Cre mRNA. Data quantified by flow cytometry 72 hours after VLP addition, n = 3 replicates.

H. Functional transfer of RNA into loxP-GFP N2a cells mediated by VSVg pseudotyped VLPs produced with rHsPeg10 and Hs.cargo(Cre) or Cre mRNA. Data quantified by flow cytometry 72 hours after VLP addition, n = 3 replicates. For all panels *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, One-Way ANOVA.