RNA Isolation and Profiling Services for Extracellular Vesicle

The profiling of RNA for EVs (Extracellular vesicles) can reveal the molecular mechanisms of cellular pathologies in which EVs are involved, facilitating the discovery of EVs-based disease biomarkers and the development of potential therapeutic strategies. Creative Biolabs has built a comprehensive RNA isolation and profiling technology platform for EVs, assisting customers in revealing the EVs-based RNA molecular cargo and its potential functions.

Abundant RNA Contents in EVs

EVs are involved in a wide variety of biological processes, from cell-cell communication to tissue repair and immunity. EVs and their RNA cargo have been extensively explored as potential biomarkers for various diseases and therapeutic targets. EVs contain RNA molecules, such as mRNAs, sRNAs, and miRNAs, that are important for cellular gene expression and protein organization. These molecules can be exploited to identify aberrantly expressed genes and pathways, diagnose differences in gene expression and understand the state of EVs-induced cytopathology. RNA isolation and analysis of EVs is a process designed to capture and isolate significant RNA molecules from EVs and a powerful tool for interrogating the molecular components of EVs, which can provide valuable data for many biological studies.

Fig.1 Principles of functional cell communication by extracellular vesicle RNA. (O'Brien, 2020)

Methods of RNA Isolation and Analysis for EVs in Creative Biolabs

In general, RNA in EVs can be extracted by cell precipitation, separation and, non-lipid filtration methods. The cell precipitation method involves placing the cell suspension in a clean centrifuge tube, followed by precipitation of the cells with specific reagents, and filtering the filtrate after cell precipitation to obtain EVs and their RNA. The separation method uses specific antibodies or specific antigens to separate the vesicles from the cells and extract RNA from the suspension, while the non-lipid filtration method is based on the property of the vesicles themselves to encapsulate RNA, and the suspension is filtered through a specific filter sieve to extract RNA from the vesicles.

RNA isolation from EVs requires breaking the membrane of EVs to determine the total RNA contents. Various enzymatic techniques are used to disrupt the surface membrane of EVs, which allows the RNA content in EVs to be distributed and collected. Once released, the RNA content can be further purified and characterized by nucleic acid isolation and analysis techniques such as PCR, RT-PCR, and RNA sequencing. The resulting data can contribute to determining the functional role of EVs in cellular processes and the complex interactions that occur between endogenous molecules and exogenous messengers. In addition, it could allow researchers to understand the molecular information network that EVs deliver to cells in order to manipulate cellular metabolism, signaling pathways, and genetic responses.

Fig.2 Graphical presentation of basic steps and possible variability factors in the EV processing and EV nucleotide sequence analysis. (Tesovnik, 2021)

Applications

RNA profiling of EVs can be applied to identify disease-associated pathways involved in EVs, infer gene collaboration, and understand EVs-mediated molecular networks that drive cellular behavior. By analyzing the content of EVs RNA, researchers can elucidate the genome-wide impact of signaling processes involved in EVs and their unique and universal responses to physiological and pathological conditions. Furthermore, it also facilitates the development of new therapeutic strategies based on EVs and the monitoring of disease treatment progression, manipulating the cellular RNA landscape and exploiting it to play an active role in maintaining homeostasis.

The extensive RNA in EVs provides a rich source of molecular information for understanding the specific properties and functions of EVs and their donors. Creative Biolabs provides high-quality RNA isolation and profiling services for EVs, allowing clients to advance further research into the molecular information of EVs. Please contact us to discuss your project. Please contact us to advance your research.

References

O'Brien, K.; et al. RNA delivery by extracellular vesicles in mammalian cells and its applications. Nat Rev Mol Cell Biol. 2020, 21(10): 585-606.
Tesovnik T, et al. Technological approaches in the analysis of extracellular vesicle nucleotide sequences. Front Bioeng Biotechnol. 2021, 9: 787551.

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