RNA sequencing is a powerful tool for studying extracellular vesicles (EVs). It can serve to characterize the composition and function of EVs, as well as identify novel biomarkers and therapeutic targets. Creative Biolabs offers RNA sequencing and profiling services for multiple types of RNA in EVs, including transcriptome, miRNA, mRNA, lncRNA, and circRNA.

RNA Sequencing Procedures for EVs

1. Isolation of EVs: EVs can be isolated from the sample using a variety of techniques, including ultracentrifugation, size-exclusion chromatography, and immunoaffinity purification.
2. Purification of RNA from EVs: RNA can be isolated from the isolated EVs using a variety of methods, including phenol-chloroform extraction and column-based purification.
3. cDNA Synthesis: cDNA can be synthesized from the purified RNA using reverse transcription.
4. Library Preparation: cDNA libraries can be prepared for sequencing, using PCR-based methods.
5. Sequencing of Libraries: The libraries can be sequenced using either next-generation sequencing or Sanger sequencing.
6. Data Analysis: The resulting sequencing data can be analyzed using bioinformatics tools to identify and quantify expressed genes.

RNA Sequencing Services for EVs at Creative Biolabs

RNA sequencing services for EVs provide important insights into the nature of RNA cargo based on the intercellular transfer of EVs, contributing to the elucidation of the mechanisms by which EVs interact with their environment as well as the molecular mechanisms associated with their potential regulatory and therapeutic effects. RNA sequencing services for EVs at Creative Biolabs include, but are not limited to:

• Whole Transcriptome Sequencing Service
• miRNA Sequencing Service
• mRNA Sequencing Service
• lncRNA Sequencing Service
• circRNA Sequencing Service

Fig.1 The distinct RNA contents for EVs. (Kim, 2017)

Application of EVs RNA sequencing

The investigation of the whole transcriptome of EVs in RNA sequencing enables the identification of genes expressed from the parental source of EVs, as well as the measuring of the abundance of specific transcripts, facilitating insight into the role played by EVs in health and disease. In addition, RNA sequencing serves to determine the amount of mRNA and non-coding RNA present in EVs, as well as to study post-transcriptional modifications of mRNA. These types of profiling can be used to identify biomarkers for diseases associated with EVs, examine the role of EVs in disease progression, and identify novel therapeutic targets and mechanisms.

Fig.2 Mechanisms of action for EV-RNA. (Cocks, 2021)

RNA sequencing for EVs supports the exploration of the molecular composition of EVs and the biological processes they are involved in, helping to better understand the mechanisms of EV-mediated communication between cells and tissues, which can be used to develop new diagnostic and therapeutic approaches. Creative Biolabs offers RNA sequencing services with an innovative molecular analysis platform for EVs. Please contact us for a customized quote.

References

1. Kim, K.M.; et al. RNA in extracellular vesicles. Wiley Interdiscip Rev RNA. 2017, 8(4): 10.1002/wrna.1413.
2. Cocks, A.; et al. Diverse roles of EV-RNA in cancer progression. Semin Cancer Biol. 2021, 75: 127-135.

• DNA Isolation and Profiling Services
• RNA Isolation and Profiling Services
• Protein Isolation and Profiling Services
• Cytokines Profiling Service