acRIP Sequencing

Acetylated RNA immunoprecipitation (acRIP) sequencing is a next generation sequencing (NGS)-based method to comprehensively detect global maps of N4-acetylcytidine (ac4C) modification, a new type of RNA modification, at high efficiency. Our experienced technical team provides you with integrated acRIP sequencing service in order to detecting and analyzing all ac4C modification in a sample, offering a powerful tool for epigenetics research.

Overview

ac4C is a conservative chemical modification in eukaryotes and prokaryotes. Earlier researches believed that ac4C mainly exists in tRNA and 18S rRNA, recent studies have shown that ac4C is also abundant in mRNA, and most of the enrichment sites appear in coding sequence (CDS). ac4C acetylation is the acetylation of N4-acetylcytosine under the action of ac4C modifying enzyme. As a new type of RNA modification, ac4C acetylation contributes to protein translation, affecting RNA stability and alternative splicing, and regulating gene expression, which is expected to become a novel approach of epigenetic transcriptomics. Our company provides comprehensive acRIP sequencing service, which can quickly identify the complete picture of ac4C modification in multiple types of RNA molecules. Based on the principle that antibody specifically binds to acetylated modified bases, the technique uses acetylated modified fragments enriched by RNA co-immunoprecipitation as raw materials, extracts mRNA containing ac4C modifications, and combines high-throughput sequencing technology with bioinformatics analysis, which facilitates systematic studies of ac4C modification in transcriptome. acRIP sequencing can comprehensively analyze the distribution and changes of ac4C in transcriptome, help researchers deeply understand the role of RNA acetylation in life and diseases, and promote precise diagnosis and treatment of diseases.

Features

High Efficiency Wide Coverage Strict Quality Control Rich Experience
Use carefully optimized experimental procedures, it has extremely high efficiency and specificity. Comprehensively detect ac4C acetylation sites of mRNA, circRNA, LncRNA, pri-miRNA, rRNA, tRNA, etc. Quality control at every stage from library construction to bioinformatics analysis. Provide professional bioinformatics analysis team to meet comprehensive needs of customers.

Project Workflow

Sample Preparation

1. Sample Preparation

RNA purification;
quality assessment and quantification.

Library Preparation

2. Library Preparation

RNA fragmentation;
immunoprecipitation;
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.

acRIP Sequencing

Bioinformatics Analysis Pipeline

In-depth data analysis:

  • Statistics of peaks distribution
  • Peaks visualization
  • Evolutionary conservative analysis
  • Motif search
  • Differential binding analysis
  • Differential binding peaks annotation
  • Enrichment analysis of peaks genes
  • Differential correlation analysis of ac4C modification

Sample Requirements

RNA sample quantity ≥ 50 ug.
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. Arango D, Sturgill D, Alhusaini N, et al. Acetylation of Cytidine in mRNA Promotes Translation Efficiency. Cell, 2018, 13, 175(7): 1872-1886.
  2. Roundtree I A, Evans M E, Pan T, et al. Dynamic RNA Modifications in Gene Expression Regulation. Cell, 2017, 169, 1187-1200.
  3. Kumbhar B V, Kamble A D, Sonawane K D. Conformational preferences of modified nucleoside N4-acetylcytidine, ac4C occur at "wobble" 34th position in the anticodon loop of tRNA. Cell Biochem Biophys, 2013, 66, 797-816.
  4. Agarwala S D, Blitzblau H G, Hochwagen A, et al. RNA methylation by the MIS complex regulates a cell fatedecision in yeast. PLOS Genet, 2012, 8: e1002732.
* For Research Use Only. Not for use in diagnostic procedures.


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