Recent years there has been a considerable increase in interest in circRNA studies, especially circRNA expression analysis. CD Genomics provides circRNA sequencing (circRNA-seq) service for identifying circRNAs, quantifying circRNA expression levels, and characterizing their potential functions. Please contact our expert team to discuss how our circRNA-seq service can refine your NGS projects.
Circular RNAs (circRNAs) are a large class of naturally occurring endogenous non-coding RNAs (ncRNAs) that contain a covalent circular structure formed by back-splicing without a 3' tail and or 5' cap. Previous studies have shown that circRNAs are widely present in a relatively low abundance compared with linear mRNA molecules and some are conserved in eukaryotic organisms. Although the function of most circRNAs is still ambiguous, studies have revealed that circRNAs function as molecular sponges to sequester specific microRNAs (miRNAs) repress their endogenous activity. Studies also highlighted the critical roles of circRNAs in cellular metabolic processes and cancer occurrence and progression. Thus, circRNA expression analyses help interpret molecular fundamentals of complex traits and diseases. RNA sequencing (RNA-seq) has emerged as a transcriptome-wide method to characterize and quantify circRNAs. A number of computational approaches have been developed for extracting, characterizing, and quantifying circRNAs. Dissection of changes in circRNA expression among different samples is a routine analysis in circRNA studies to evaluate the significance of differentially expressed circRNAs. CD Genomics offers circRNA-seq to investigate miRNA sponges and the regulatory function in gene expression.
|The flexibility to prepare libraries and analyze data according to your needs.||Extensive experience in dealing with numerical samples and data analysis.||Generating high quality data with a guaranteed Q30≥ 80%.||Employing Industry standard software and in-house bioinformatics pipelines.|
RNA purification; quality assessment and quantification
Ribosomal RNA Removal
250~300 bp Insert cDNA Library
Illumina Novaseq, PE 150
≥ 40 million read pair per sample
Visualize and preprocess results, and perform custom bioinformatics analysis.
RNA sample (quantity ≥ 5 μg, concentration ≥ 300 ng/μL)
RIN ≥ 7.0, with smooth baseline; OD260/280: 1.8-2.2; OD260/230: ≥ 1.8.
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: raw data as BAM files, coverage summary, QC report, custom bioinformatics analyses.
Pie chart of circRNA type
CircRNA expression density
Differential circRNA expression volcano plot
circRNA cluster analysis
KEGG pathway annotation
Intrahepatic cholangiocarcinoma (ICC) is a primary liver cancer, accounting for approximately 10-20% of all primary liver cancers and exhibiting a rapid increase in incidence over the past few decades. Understanding the pathogenic mechanisms underlying ICC is crucial for the development of effective treatment strategies. Circular RNA (circRNA) is a class of non-coding RNA known for its diverse functions, including miRNA sponging and regulation of gene expression. The role of circRNAs in ICC progression remained an area of active research.
CircACTN4 Overexpression in ICC Tissues
CircACTN4 Promotes ICC Cell Proliferation and Invasion
CircACTN4 Regulates FZD7 Expression in ICC Cells
Interaction Between CircACTN4 and YBX1
CircACTN4 Recruits YBX1 to Activate FZD7 Transcription
CircACTN4 Upregulates YAP1 Expression by Sponging miR-424-5p
CircACTN4 Involvement in Wnt/Hippo Signaling
We recommend a minimum of 5 µg of Total RNA for each submission. For detailed submission guidelines, please refer to our Sample Submission Recommendations.
It is generally recommended to generate at least 10 GB of sequencing data to ensure sufficient coverage for circRNA analysis, based on saturation analysis.
Yes, it is essential. We recommend a minimum of three biological replicates, although more replicates are beneficial for robust results.
In lncRNA data (10 GB clean data), typically, you can identify around 1,000 to 2,000 circRNAs. In contrast, the Rnase enrichment method can identify 20,000 to 30,000 circRNAs, with data quantities in the tens of thousands. We recommend these approaches based on your research goals. If your study is exploratory, lncRNA library sequencing offers cost-effectiveness. For targeted circRNA research, the standard circRNA library preparation method is recommended.
CircRNA can be validated through two methods:
CircRNAs are known for their resistance to RNase R degradation. However, degradation can occur through two processes: RNase R degradation and hydrolysis. Hydrolysis affects both circular and linear RNA and can occur more readily in circular RNA due to the close proximity of base pairs. To maintain circRNA stability, they should be stored at low temperatures and away from magnesium ions. While RNase R degradation can enhance sensitivity in circRNA detection, it may not improve specificity.
We recommend using sequencing technology for circRNA analysis as it enables the discovery and identification of new results.
Yes, circRNA levels can vary significantly between samples. Unlike most genes, circRNAs are not part of the "housekeeping" genes and should not be normalized in the same manner. Their expression levels can exhibit substantial differences between samples.
Yes, circRNA research can benefit from combining with other sequencing techniques. For example, you can study the interaction of specific RNA-binding proteins (RBPs) with circRNAs using RIP-seq. At the genome level, ATAC-seq and ChIP-seq can help explore the transcriptional activity and regulatory elements influencing circRNAs. These approaches can also assess whether linear host RNA and circRNA share similar transcriptional regulation.