Exosomal RNA sequencing has become a valuable tool in molecular biology, offering a detailed look at the mechanisms of intercellular communication. By analyzing the RNA cargo carried by exosomes, this technique provides important insights into their roles in normal physiological functions and disease-related processes.
Image Credit: Norgen Biotek Corp.
This article examines the importance and relevance of exosomal RNA sequencing, looking at its principles, applications, and impact on disease research, biomarker discovery, and clinical diagnostics.
Understanding exosomes and their RNA cargo
Knowing the role of exosomes in the human body is important to understanding the impact of exosomal RNA sequencing.
Exosomes are small, extracellular vesicles that are released by cells into various bodily fluids, including blood, urine, and cerebrospinal fluid. These vesicles support intercellular communication, enabling the transfer of RNA and other bioactive molecules between cells.
The workflow of exosomal RNA sequencing
Exosomal RNA sequencing unravels the RNA content of exosomes via a series of specific steps. This workflow traditionally features exosome isolation, RNA purification and isolation, library preparation, sequencing, and bioinformatics analysis.
Exosomes are initially isolated from the desired biological sample, usually via size-based isolation, differential ultracentrifugation, or polymer-based precipitation. RNA is then extracted from exosomes via specialized protocols designed to maintain the RNA integrity, for example, silica or silicon carbide-based spin column technology.
After the RNA is extracted, it undergoes library preparation. Reverse transcription is used to generate complementary DNA (cDNA) from the RNA before the cDNA is subjected to high-throughput sequencing technologies such as next-generation sequencing (NGS).
This enables the acquisition of vast amounts of sequencing data, which is processed via bioinformatics analysis. Analysis steps include read alignment, identification of RNA species, quantification of expression levels, and interpretation of downstream data.
Image Credit: Norgen Biotek Corp.
Applications of exosomal RNA sequencing
Exosomal RNA sequencing offers potential benefits to a wide range of applications, considerably advancing basic research and clinical diagnostics.
Biomarker discovery
The discovery of exosomal RNA biomarkers is one clinically significant application of RNA sequencing.1,2 Researchers can investigate the RNA profiles of exosomes derived from biofluids, such as blood or urine, to identify specific RNA molecules that can function as biomarkers for different diseases.
These biomarkers offer significant benefits to disease diagnosis, prognosis, and monitoring, as well as offering non-invasive alternatives to conventional tissue biopsies.
Cancer research
Exosomal RNA sequencing is an essential component of cancer research.3
Analyzing cancer-derived exosomes’ RNA content affords researchers insight into tumor-specific gene expression patterns, allowing researchers to identify potential therapeutic targets and understand metastasis and drug resistance mechanisms.
Exosomal RNA sequencing also facilitates liquid biopsies, enabling the non-invasive detection and monitoring of cancer by analyzing circulating exosomal RNA.
Disease pathogenesis
Studying the RNA cargo of exosomes also enables a deeper understanding of disease pathogenesis, with exosomal RNA sequencing helping to reveal the role of exosomal RNA in disease progression, tissue regeneration, immune responses, and a range of neurological disorders.3
Identifying disease-specific RNA signatures within exosomes allows researchers to gain useful insights into disease mechanisms, leading to the development of novel therapeutic strategies.
Drug development and treatment monitoring
Exosomal RNA sequencing supports drug development and treatment monitoring.4
The assessment of changes in exosomal RNA profiles both before and following treatment supports the evaluation of treatment responses, the identification of drug resistance mechanisms, and the discovery of potential therapeutic targets.
This information can be used to underpin personalized medicine approaches, enabling tailored treatments based on individual patients’ RNA signatures.
Image Credit: Norgen Biotek Corp.
Norgen Biotek: A pioneer in exosome research
Norgen Biotek is at the forefront of exosome research, offering dependable, all-in-one solutions for the exosome research pathway.
A range of exosome purification kits is available, tailored for a diverse array of biofluids and input volumes. Each kit features a highly selective, proprietary silicon carbide resin matrix designed to isolate exosomes while ensuring optimal purity.
Norgen Biotek’s RNA extraction kits can be used following exosome isolation, enabling the isolation of high-quality exosomal RNA. Optimized protocols ensure minimal RNA degradation while recovering all sizes of RNA and ensuring this is suitable for sensitive downstream applications such as next-generation sequencing.
Norgen Biotek also offers comprehensive solutions for exosomal RNA library preparation and sequencing, with library preparation kits designed to enable the conversion of exosomal RNA into complementary DNA (cDNA) – a critical pre-sequencing step and key to ensuring the generation of high-quality sequencing libraries.
Support is also available beyond sample preparation and sequencing, and Norgen Biotek’s bioinformatics experts can provide tailored analysis pipelines for exosomal RNA sequencing data. These bioinformatics solutions provide valuable insights into the functional annotation and interpretation of their data, from read alignment to the identification of RNA species and expression quantification.
Norgen Biotek’s commitment to advancing exosome research is clear, as evidenced by its comprehensive portfolio of exosome purification and exosomal RNA isolation solutions and its downstream applications, such as RNA sequencing.
Norgen Biotek empowers researchers by providing reliable products, sequencing services, and bioinformatics support.
References and further reading
- Boukouris, S. and Mathivanan, S. (2015). Exosomes in bodily fluids are a highly stable resource of disease biomarkers. PROTEOMICS - Clinical Applications, 9(3-4), pp.358–367. https://doi.org/10.1002/prca.201400114.
- Elkommos-Zakhary, M., Rajesh, N. and Beljanski, V. (2022). Exosome RNA Sequencing as a Tool in the Search for Cancer Biomarkers. Non-Coding RNA, (online) 8(6), p.75. https://doi.org/10.3390/ncrna8060075.
- Zhu, L., et al. (2020). Isolation and characterization of exosomes for cancer research. Journal of Hematology & Oncology, (online) 13, p.152. https://doi.org/10.1186/s13045-020-00987-y.
- Stevic, I., Buescher, G. and Ricklefs, F.L. (2020). Monitoring Therapy Efficiency in Cancer through Extracellular Vesicles. Cells, 9(1), pp.130–130. https://doi.org/10.3390/cells9010130.
Acknowledgments
Produced from materials originally authored by Rahaf Alhadadin from Norgen Biotek Corporation.
About Norgen Biotek Corp.
Norgen Biotek: Advancing science with best-in-class, scientist-backed innovations
Norgen Biotek is a fully integrated biotechnology company that focuses on providing complete workflows for molecular biology sample preparation and analysis. With a diverse portfolio of over 600 products, the company delivers high-performance, user-friendly, and cost-effective solutions.
Scientifically driven, industry trusted
Norgen kits cover a broad range of applications from collection and preservation to isolation and purification. Our expert R&D team continuously develops cutting-edge technologies that set new industry standards for RNA, DNA, protein, and exosomal isolation, ensuring superior yield, purity, and integrity from even the most challenging sample types.
Unparalleled performance
At the heart of Norgen’s success is its patented Silicon Carbide (SiC) Technology. This proprietary resin exhibits uniform binding affinity for all RNA species, regardless of molecular weight or GC content. This ensures the full diversity of small and microRNA are captured while eliminating the need for phenol extraction. This innovative technology sets Norgen kits apart from others, positioning them as leaders in RNA purification.
Comprehensive solutions for any challenge
Norgen is committed to providing high-quality kits capable of processing a wide range of sample types, from ultra-low input samples such as liquid biopsies to highly impure samples like stool or soil. Our sample collection and preservation devices for stool and saliva simplify handling by rendering samples non-infectious by preventing microbial growth and inactivating viruses, while our blood and urine preservation solutions ensure the stability of highly vulnerable cell-free nucleic acids.
To meet varying research demands, we offer multiple isolation methods including, but not limited to high-throughput and automation-ready magnetic bead-based formats. Additionally, our multiple-analyte kits enable the simultaneous purification of RNA, DNA, and proteins, maximizing data extraction from a single sample.
Norgen offers an extensive variety of TaqMan qPCR kits designed for molecular diagnostic use, including lyophilized kits for easy shipping. Library preparation kits for both DNA and RNA samples are also available to support genomic applications. Norgen recently released their EXTRAClean technology, an innovative solution that minimizes background noise while providing high-purity RNA, significantly enhancing NGS performance.
Why choose Norgen?
- Scientist-Driven Innovation – Developed by leading experts in molecular biology
- Proven Quality & Reliability – ISO 9001 and ISO 13485 certified, indicating a commitment to selling high-quality products
- Global Presence – We ship to over 150 countries and have a network of 60+ distributors
- Award Winning – Norgen Biotek Corp. was honored with the 2021 Innovative Leaders Award, and recognized as the 2024 Rapid Star Award Winner in the PCR category.
- Innovative Products – Hold more than thirty issued and pending patents for products presenting solutions for all research & clinical applications
Driven by a mission to accelerate scientific discoveries, Norgen Biotek actively supports researchers by providing educational resources, technical workshops, and application notes. Their NorBlog serves as a hub for the latest scientific discoveries, protocol optimizations, and industry trends.
Explore Norgen Biotek’s innovative solutions today and take your research to the next level.
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