Exosomal RNA Sequencing

Exosomes are tiny membrane-bound extracellular vesicles produced in the endosomal compartment of most eukaryotic cells and can be found in abundance in many and perhaps all body fluids including blood, urine, cerebrospinal fluid. These tiny vesicles contain distinct subsets of proteins, lipids, DNA, and RNA (including microRNAs, mRNAs, lncRNAs and other ncRNAs). Exosomes have a unique biogenesis pathway via multivesicular bodies and may play an important role in cell-to-cell signaling, thereby influencing the immune system and mediating adaptive immune responses to pathogens and tumors. The delivery of their cargo RNA molecules is a mainstream hypothesis to explain biological effects.

Overview

Exosomes contain RNA, proteins, lipids, and metabolites that reflect the type of cell origin. Thus, proteomics, transcriptomics, and metabonomics are often performed for large-scale analyses of exosomes. With the advent of next-generation sequencing technology, exosome research is accelerating not only in cancer, but also in various diseases. It has been found that exosomes, based on their specific membrane and cytoplasmic proteins, as well as other protein subsets, may be associated with different types of cell-related functions and may also be associated with human diseases. Knowledge of exosome RNA sequencing not only helps to understand their biological roles, but analysis of exosome RNA-Seq data based on bioinformatics can provide a new way to search for new disease-related biomarkers in the body fluids of patients. CD Genomics provides high-throughput sequencing services to quickly and efficiently obtain the global information of exosomal RNAs. Our one-step exosomal RNA sequencing services cover the detailed analysis of all types of exosomal RNA, including exosomal small RNA (mainly exosomal microRNA), exosomal lncRNA, exosomal circRNA, and exosomal mRNA).

Service Portfolio

Explore how biofluid profiling using NGS helps researchers understand dynamics and interactions of exosomal RNA.

Exosomal microRNA Sequencing

Our exosomal microRNA sequencing examines monitor global microRNA expression at an affordable price, enabling the identification of biomarkers associated with diseases like cancer.

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Exosomal Small RNA-Seq

Exosomal small RNA sequencing is a powerful tool for analyzing small RNAs including miRNAs, piRNAs and siRNAs, offering both quantitative and qualitative information.

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Exosomal lncRNA-Seq

Exosomal lncRNA sequencing examines global lncRNAs in exosomes. As exosomal lncRNAs are involved with tumorigenesis, tumor angiogenesis, and chemoresistance, this service can detect promising biomarkers for cancer.

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Exosomal Long RNA-Seq

Exosomal long RNA-seq analyzes mRNAs, long non-coding RNAs, and circular RNAs in the sample, enabling alternative splicing analysis and detection of novel transcripts, and gene fusion events.

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Exosomal mRNA-Seq

Exosomal mRNA sequencing service can not only help you to profile exosomal mRNA with regard to the expression levels and dynamics but also understand the physiological roles, with or without knowledge of priori sequences.

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Exosomal circRNA-Seq

Exosomal cicrRNA-Seq can quickly and efficiently obtain global information on exosomal circRNAs. Our single-base resolution technology allows the detection of circRNAs from very small amounts of cellular material.

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Features

Superb Expertise Transcriptome-Wide Accurate Annotation One-Stop Solution
Decade of experience in high-throughput sequencing and bioinformatics analysis. Profile global exosomal RNA, including small RNA, microRNA, and lncRNA. Provide complete annotate information with multiple databases for exosomes or vesicles. One-stop solution from sample QC and library construction, to sequencing and data analysis.

Project Workflow

Sample Preparation

1. Sample Preparation

RNA purification;
quality assessment and quantification

Library Preparation

2. Library Preparation

RNA fragmentation;
m7G/m3C library preparation.

Sequencing

3. Sequencing

Illumina HiSeq;
PE 50/75/100/150.

Data Analysis

4. Data Analysis

Visualize and preprocess results, and perform custom bioinformatics analysis.

Bioinformatics Analysis Pipeline

Bioinformatics Analysis Pipeline

In-depth data analysis:

  • Length distribution and quantitative analysis of exosomal RNA
  • Differential expression analysis of exosomal RNA
  • Prediction of exosomal RNA and binding sites
  • GO annotation analysis
  • KEGG pathway analysis
  • Interactive network analysis
  • Coding-non-coding gene co-expression network (CNC network)
  • lncRNA-miRNA-mRNA network

Sample Requirements

RNA sample (concentration ≥ 1 ng/uL, quantity ≥ 2 ug)

1.8 ≤ OD260/280 ≤ 2.2, OD260/230≥2.0, RIN ≥ 6.5, 28S:18S ≥ 1.0.

Please make sure that the RNA is not significantly degraded.

Sample storage: RNA can be dissolved in ethanol or RNA-free ultra-pure water and stored at -80°C. RNA should avoid repeated freezing and thawing.

Shipping Method: When shipping RNA samples, the RNA sample is stored in a 1.5 mL Eppendorf tube, sealed with sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.

Deliverable: FastQ, BAM, coverage summary, QC report, custom bioinformatics analysis.

References:

  1. Huang X, Yuan T, Tschannen M, et al. Characterization of human plasma-derived exosomal RNAs by deep sequencing. BMC genomics. 2013, 14(1):319.
  2. Raimondo F, Morosi L, Chinello C, et al. Advances in membranous vesicle and exosome proteomics improving biological understanding and biomarker discovery. Proteomics. 2011, 11(4):709-20.
* For Research Use Only. Not for use in diagnostic procedures.


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