Exosomal circRNA Sequencing

Circular RNAs (circRNAs) are non-coding endogenous RNAs discovered in all eukaryotic cells. These molecules are involved in multiple cellular functions, which may include acting as modulators of gene expression, affecting biological processes (such as cell proliferation and invasion). Research has shown that circRNA was also detected in small extracellular vesicles called exosomes, which ubiquitously exit in many and perhaps all body fluids including blood, urine, cerebrospinal fluid. It's promising to discriminate patients from healthy individuals, identifying new potential exosome-based disease especially tumor biomarkers by investigating the presence and expression levels of exosomal circRNAs.

Overview

circRNAs are single-stranded cyclized molecules that are usually produced during the process of pre-mRNA "back splicing". Unlike linear RNAs, circRNAs possess a special structure with certain properties. circRNAs lack a poly(A) tail, which makes them stable and resistant to exonucleases (including RNase R), and make them have a median half-life 2.5 times that of their linear counterpart. Studies reported that circRNAs are enriched in exosomes, and play a pivotal role in cancer biology.

Advances in large-scale next-generation sequencing technologies have increased the understanding of exosomal RNA biology in an exponential manner. CD Genomics offers high-throughput sequencing services to 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. Moreover, exosomal circRNA sequencing services can help you not only analyze exosomal circRNAs based on expression levels and dynamics, but also understand their biological functions and clinical significance, thus elucidating the relationship between circRNAs and human diseases, including neurological disorders, cardiovascular diseases, metabolic diseases, and tumors.

Features

Superb Expertise Transcriptome-Wide Bioinformatics Analysis One-Stop Solution
Decade of experience in high-throughput sequencing and bioinformatics analysis. Accurately profile the presence and expression levels of exosomal circRNAs of transcriptome-wide. Our integrated bioinformatics pipeline can be tailored to suit your project. One-stop solution from sample handling to detailed interpretation.

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 circRNA
  • Differential expression analysis of exosomal circRNA
  • Function annotation
  • Structure identification
  • Gene expression analysis
  • GO/KEGG analysis
  • Interactive network analysis
  • Coding-non-coding gene co-expression network (CNC 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. Fanale D, Taverna S, Russo A, Bazan V. Circular RNA in exosomes. InCircular RNAs. 2018 (pp. 109-117).
  2. Shi X, Wang B, Feng X, et al. circRNAs and exosomes: a mysterious frontier for human cancer. Molecular Therapy-Nucleic Acids. 2020, 19:384-92.
* For Research Use Only. Not for use in diagnostic procedures.


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