Exosome Marker Lentiviral Particles Facilitate Exosome Tracking and Biodistribution Studies

Exosomes have many biological functions as short- and long distance nanocarriers for cell-to-cell communication. They allow the exchange of complex information between cells, and thereby modulate various processes such as homeostasis, immune response and angiogenesis, in both physiological and pathological conditions. In addition, due to their unique abilities of migration, targeting, and selective internalization into specific cells, they are promising delivery vectors.

Exosome Research Challenges

Exosomes offer a potential new frontier for diagnosis and therapy and may serve as an alternative to cell-based therapeutic approaches. However, a major drawback for translating exosome treatment to the clinic is that current understanding of these endogenous vesicles is insufficient, especially in regards to their in vivo behavior. Tracking exosomes in vivo can provide important knowledge regarding their biodistribution, migration abilities, toxicity, biological role, communication capabilities, and mechanism of action. Therefore, the development of efficient, sensitive and biocompatible exosome labeling and imaging techniques is highly desired.

Elucidating strategies for in vivo exosome transport and selective targeting. Fig. 1 Elucidating strategies for in vivo exosome transport and selective targeting. (Betzer, et al., 2020)

Exosome Marker Lentiviral Particles

Recent studies have developed different methods for exosome labeling and imaging, which have allowed for in vivo investigation of their bio-distribution, physiological functions, migration, and targeting mechanisms. Among them, the technique of fusion expression of fluorescent proteins (e.g., green fluorescent protein, GFP) with marker proteins such as CD63, CD9, and CD81 on the exosomal membrane using molecular biology is considered to be an effective approach.

Creative Biogene has developed a series of ready-to-use exosome marker lentiviral particles by constructing plasmids of the exosome marker proteins CD63, CD9, CD81 and expression elements of different fluorescent markers (tdTomato, mCherry, GFP) and then packaging them into lentiviruses. Infection of cells with these recombinant lentiviral particles allows the cells to secrete exosomes with fluorescence.

Exosome Marker Lentiviral Particles
EF1a-CD9-tdTomato Lentivirus	EF1a-CD63-tdTomato Lentivirus
EF1a-CD81-tdTomato Lentivirus	EF1a-FLAG-mCherry-CD63 Lentivirus
EF1a-FLAG-Luc-CD63 Lentivirus	EF1a-Halo-CD63 Lentivirus
EF1a-Avi-CD63 Lentivirus	CMV-CD63-GFP-FLAG Lentivirus
CMV-CD81-GFP-FLAG Lentivirus	CMV-CD63-mCherry-FLAG Lentivirus
CMV-CD81-mCherry-FLAG Lentivirus	CMV-CD9-mCherry-FLAG Lentivirus
CMV-FLAG-GFP-CD63 Lentivirus

Advantages of Exosome Marker Lentiviral Particles

Exosome marker lentiviral particles are a convenient-to-use tool to track the movement of exosomes from donor to recipient cells. By monitoring the fluorescence of membrane proteins, direct visualization of exosome transfer in live cultures and in vivo can be achieved. In addition, this labeling strategy avoids the false positive signals of labeling other lipid entities because fluorescent proteins are specifically affixed to exosomes.

Applications of Exosome Marker Lentiviral Particles

Applications

Exosome quantitation
Exosome biodistribution studies
Exosome animal study tracking
Cell uptake studies
Cell targeting studies
Drug library screening

Application Cases

Live Tracking of Inter-organ Communication by Endogenous Exosomes In Vivo

Fig. 2 Identification and in vivo visualization of EVs in zebrafish. (Verweij, et al., 2019)

Researchers express the CD63-pHluorin reporter gene in zebrafish embryos for tracing exosomes in vivo. This fluorescent reporter molecule specifically targets (late) endosomes and is secreted on exosomes, allowing visualization of exosome release from individual cells in vitro. The combination of imaging methods and label-free proteomic quantification allows tracking throughout the animal until it reaches its final destination and reveals its biological function. The study revealed a massive release of exosomes from the yolk syncytium layer (YSL) into the blood stream, which are captured, endocytosed and degraded by macrophages and endothelial cells in the caudal vein plexus with scavenger receptor and viability-dependent properties. In conclusion, this study reveals for the first time the secretion, diffusion and uptake of endogenous exosomes by target cells in vivo and the key molecular mediators of these processes.

References:

Betzer, O.; et al. Advances in imaging strategies for in vivo tracking of exosomes. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2020 Mar;12(2): e1594.
Verweij, F. J.; et al. Live Tracking of Inter-organ Communication by Endogenous Exosomes In Vivo. Dev Cell. 2019 Feb 25;48(4):573-589.e4.

* For research use only. Not intended for any clinical use.

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Exosome Marker Lentiviral Particles Facilitate Exosome Tracking and Biodistribution Studies

Exosome Research Challenges

Exosome Marker Lentiviral Particles

Advantages of Exosome Marker Lentiviral Particles

Applications of Exosome Marker Lentiviral Particles

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