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EV Shuttles and EV-Entry System

Next generation transfection
system using exosomes

Exosomes as transfection shuttles

  • Delivery to transfection-resistant cells
  • Simple loading protocol
  • Non-viral delivery platform
  • Create stable cell lines using EV shuttles
  • Boost exosome delivery with EV-Entry

The EV-Entry System

   Labeled exosome protein delivery

Extracellular vesicles (EVs), including exosomes, are nanocarriers used by cells to transport RNA and protein signals and are central to intercellular communication. EVs can be added to recipient cells to deliver exogenous cargo. The EV-Entry reagent system not only enhances the rate of uptake of EVs by the recipient cells but also increases the offloading of the transported cargo into the cytoplasm of the recipient cells. EV-Entry boosts the delivery of both RNA and protein cargo. The EV-Entry system is comprised of Reagent A and B components that are mixed together with EVs or exosomes just prior to use. The protocol takes less than an hour and highly efficient at enabling EVs and exosomes to deliver cargo to recipient cells.

Sample Enhanced Exosome Cargo Delivery with EV-Entry

Exosomes were labeled with ExoGreen (cat# EXOG200A-1), a dye that binds to exosomal proteins and added on HEK cells with or without the EV-Entry reagent. Cells were imaged 18h post incubation of labeled exosomes with the cells. Cells that received exosomes with the EV-Entry reagent showed remarkably enhanced uptake of exosomes as well as release of exosomal cargo in the cytoplasm as demonstrated by the staining pattern of exosomal proteins in the recipient cells using a very brief exposure time.

   XPAK-Luc exosome reporter protein delivery

XPack-Luciferase Reporter EV-Entry Data

HEK 293 cells were transfected with XPack-Luciferase (cat# XPAK732PA-1). Exosomes from transfected cells were harvested and added on to naive HEK 293 cells with or without the EV-Entry reagent. After 18h incubation, target cells were washed multiple times with PBS and lysed. Luciferase activity in the recipient cells was measured by luciferase assay. Cells that received XPack-Luciferase exosomes with the EV-Entry reagent demonstrated greater than 11-fold greater luciferase activity compared to untreated exosomes suggesting enhanced delivery of the enzyme to recipient cells with EV-Entry.

Exosome RNA EV-Entry Delivery Data

HEK EV Shuttles (100 ug) were Exo-Fected with Texas-red labeled control siRNA (50 pmol) and incubated with or without the EV-Entry reagent on HEK293 cells. Recipient cells were imaged 18 hours post incubation. The EV-Entry reagent greatly increased the exosome-mediated siRNA delivery to recipient cells.

EV-Entry RNA delivery data