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CircRNA Sequencing

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To support the recent emerging of circular RNA (circRNA) research, CD Genomics is providing the circRNA sequencing service that utilizes the latest Illumina platforms for a fast, economical, and accurate characterization.

The Introduction of CircRNA Sequencing

CircRNA is a novel member of the non-coding RNAs (ncRNAs), generated by non-sequential backsplicing of exons, introns or both. They are characterized by covalently closed loop feature thus lacking 5’ end caps and 3’ poly-A tails. CircRNAs are highly stable forms of ncRNAs due to their nuclease resistance properties. When first identified in the 1970s, circRNAs were regarded as viral genomes or byproducts of mis-splicing events. But the recent researches have revealed that circRNAs are evolutionarily conserved across plants, animals, and human beings, and that circRNAs have important biological functions.

CircRNAs may act as miRNA sponges and fulfill a regulatory function in gene expression. Remarkably, a wide range of circRNAs are abnormally expressed in specific disease contexts, which suggests their association with the occurrence and development of human diseases. CircRNAs also perform multiple functions in cellular processes including templates for translation, regulation of alternative splicing and gene expression, scaffolds for the assembly of protein complexes, and modulator of rRNA and tRNA biogenesis. Additionally, they can be used to boost immune activation for antiviral therapeutic purposes due to their intervention in immune regulation and viral infection.

Comprehensive detection of circRNAs from high-throughput transcriptome data is an initial and crucial step to study their biogenesis and function. The high-throughput sequencing of rRNA/linear RNA-depletedRNA in combination with computational tools has led to the identification of thousands of new circRNAs and analysis of their linear host transcripts in a quantitative manner in diverse organisms. Unlike miRNAs and other small RNAs, circRNAs are not easily separated from other RNA species by size or electrophoretic mobility. Consequently, they are generally retained in rRNA-depleted libraries and enriched in libraries treated with RNase R that only digest linear RNA.

Advantages of CircRNA Sequencing

  • Identifies known and novel circRNAs
  • Allows profiling of circRNAs across a wide dynamic ranges
  • Explores novel biomarkers and circRNAs regulatory networks

CircRNA Sequencing Workflow

The general workflow for circRNA sequencing is outlined below. To construct circRNA sequencing library, the first step is to deplete rRNA, followed by linear RNA digestion and strand-specific library preparation. Our highly experienced expert team executes quality management, following every procedure to ensure confident and unbiased results.

Service Specification

Sample requirements and preparation
  • Sample type: Total RNA without degradation or DNA contamination
  • Suggested amount of total RNA ≥ 5 µg
  • Sample concentration ≥ 100 ng/µl
  • Sample purity: OD260/280 = 1.8~2.2
  • All RNA samples are validated for purity and quantity
Sequencing
  • Strand-specific library preparation
  • Illumina HiSeq PE150
  • More than 80% of bases with a ≥Q30 quality score
Bioinformatics Analysis
We provide customized bioinformatics analysis including:
  • Raw data quality control
  • Reference-based mapping
  • CircRNA identification and annotation
  • Known and novel circRNA expression analysis
  • Differential expression analysis, enrichment analysis, and functional analysis
  • Origin analysis of circRNA including target miRNA prediction and circRNA-miRNA interaction network analysis

Supported by our experienced scientists and advanced technology, CD genomics can help you to acquire the circRNA sequence information with single-base resolution at one time through the high-throughput sequencing by strict quality control and advanced bioinformatics analyses. If you have additional requirements or questions, please feel free to contact us.

Reference:
Wang M, Yu F, Wu W, et al. Circular RNAs: A novel type of non-coding RNA and their potential implications in antiviral immunity. International journal of biological sciences, 2017, 13(12): 1497.

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45-1 Ramsey Road, Shirley, NY 11967, USA
Tel: 1-631-275-3058
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Email: info@cd-genomics.com