mRNA-Encapsulating Exosome Production
Since exosomes have a nature of advantages, such as their favorable pharmacokinetic and immunological properties, and excellent permeation into physiological barriers (including the blood-brain barrier, BBB). Exosomes are emerging as promising carriers for mRNA drug delivery and attracted increasing attention. To promote the application of exosomes as carriers for therapeutically related mRNA delivery, Creative Biogene are committed to offering customized services of mRNA loading into exosomes. With years of experience in exosome technology, we are confident to provide high-quality mRNA-encapsulating exosomes for global clients.
Fig 1. mRNA-encapsulated exosomes. (Aslan, C., et al. 2021)
The most used methods for mRNA loading into exosomes
So far, mRNA loading into exosomes remains a challenge, although there are several methods developed for the improvements of mRNA loading into exosomes. Here, we list some methods for encapsulating mRNA molecules in exosomes.
EP is a powerful tool to introduce genes easily into eukaryotic cells with high efficiency and low toxicity. With years of optimizing and adapting, EP is a broadly applicable and well-established method for transient mRNA transfection in a variety of different cell types, including dendritic cells (DCs), B cells, T cells, adult stem cells. Similar to the process that happens to a cell, mRNA can be entrapped in the exosomes by using EP. To perform EP, the exosomes need to be diluted in a special buffer and further purified. So far, electroporation (EP) has been mainly utilized to load small RNAs (such as miRNA and siRNA) into exosomes.
- Active loading
With the help of helper proteins, mRNA can be encapsulated in the exosomes in a more efficient manner. This method is based on the ability of some proteins to bind to specific RNA sequences.
- Exosome-liposome hybrids
It is well known as nucleic acids are easily loaded into liposomes. However, the cell transfection efficiency of liposomes is low. Since exosomes are very efficient in transferring their cargo to the target cells due to their special transmembrane proteins. So far, the combination of liposomes and exosomes is a promising strategy to efficiently deliver mRNA to a cell, although this approach has not been applied to load mRNAs. Theoretically, to produce exosome-liposome hybrids, mRNAs are firstly bound with the liposomes, then the mixture is incubated with the exosomes, followed by a separation step based on ultrafiltration.
Fig 2. Schematic of method used to engineer the exosome–liposome hybrids. (Sato, Y. T., et al, 2016)
- Transfection of donor cells
Based on the exosome-producing cell itself, large amounts of exosomes containing the desired mRNA can be obtained. This is the latest as well as the most convenient method of packaging of mRNA inside exosomes. Generally, after transfection of the maternal cells with DNA encoding the mRNA, the culture medium containing exosomes released from the cells is collected and the exosomal RNAs are identified.
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- Sato, Y. T., et al. (2016). "Engineering hybrid exosomes by membrane fusion with liposomes." Scientific reports, 6(1), 1-11.
- Aslan, C.,et al. (2021). "Exosomes for mRNA delivery: a novel biotherapeutic strategy with hurdles and hope." BMC biotechnology, 21(1), 1-12.