acRIP-seq Service

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acRIP-seq Service

IntegrateRNA offers a transcriptome-wide, efficient and unbiased ac4C detection and location service using ac4C-specific RNA immunoprecipitation with high-throughput sequencing (acRIP-seq) method. Based on advanced NGS platform and experienced bioinformatics professional, we are dedicated to provide a one-stop solution from sample to data.

Chemical modifications of RNA have emerged as a new mechanism for the post-transcriptional regulation of gene expression. Epitranscriptomic modifications, including m6A, m6Am, m1A, pseudouridine, inosine, m5C, m7G and ac4C, have the potential to control all steps of mRNA metabolism including structure, stability, location and translation. One of these modification, N4-acetylcytidine (ac4C), is an ancient and highly conserved RNA modification that is present on tRNA/rRNA and has recently been investigated in eukaryotic mRNA. ac4C helps to correctly read codons during translation and improves translation efficiency and the stability of mRNA. Moreover, recent studies have indicated that ac4C is associated with the development, progression, and prognosis of a variety of human diseases. Thus, a high resolution mapping of ac4C in the transcriptome will help us explore the mechanism by which ac4C triggers the corresponding biological effects.

Schematic diagram of ac4C detection by acRIP-seqFigure 1. Schematic diagram of ac4C detection by acRIP-seq

Workflow of acRIP-seq

  • Poly(A) RNA purification
  • RNA fragmentation (100-200 nucleotides)
  • RNA immunoprecipitation using anti-ac4C antibodies or Isotypic IgG control
  • Acetylated RNA elution and purification
  • rRNA depletion
  • In vitro transcription of acetylated RNA
  • Library preparation and high-throughput sequencing

Bioinformatics Analysis

  • Genome mapping and functional annotation of all mapped reads
  • ac4C peak calling and visualization
  • Motif analysis
  • Differential acetylation analysis
  • GO/KEGG enrichment analysis of differential gene/peak
  • Custom-tailored data analysis
    ➢ Correlation analysis between differential ac4C and differential gene
    ➢ Correlation analysis between mRNA alternative splicing and differential ac4C
    ➢ Correlation analysis between Ribo-seq and acRIP-seq

Benefits and Features

  • Generate thousands of ac4C-enriched transcribed regions
  • High resolution transcriptome mapping of ac4C
  • Cost-effective and fast turnaround time

IntegrateRNA is devoted to provide clients with high-quality data for epitranscriptome research. Through a consulting, flexible, integrated and interdisciplinary approach, we provide advice, answers and services to advance your project progress. For further information, please feel free to contact us.

Reference:

  1. Arango, D. et al. (2019). Immunoprecipitation and sequencing of acetylated RNA. BIO-PROTOCOL, 9(12).
For research use only. Not intended for any clinical use.
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