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Updated: Oct 10, 2025
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Published on: March 17, 2019
N6-Methyladenosine Methylation Analysis of Long Noncoding RNAs and mRNAs in IPEC-J2 Cells Treated With Clostridium
Jiaojiao Yang1, Qiaoli Yang1, Juanli Zhang1
1College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China.
Background:
The n6-methyladenosine (m6A) modification is present widely in mRNAs and long non-coding RNAs (lncRNAs), and is related to the occurrence and development of certain diseases. However, the role of m6A methylation in Clostridium perfringens type C infectious diarrhea remains unclear.
Methods:
Here, we treated intestinal porcine jejunum epithelial cells (IPEC-J2 cells) with Clostridium perfringens beta2 (CPB2) toxin to construct an in vitro model of Clostridium perfringens type C (C. perfringens type C) infectious diarrhea, and then used methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) to identify the methylation profiles of mRNAs and lncRNAs in IPEC-J2 cells.
Results:
We identified 6,413 peaks, representing 5,825 m6A-modified mRNAs and 433 modified lncRNAs, of which 4,356 m6A modified mRNAs and 221 m6A modified lncRNAs were significantly differential expressed between the control group and CPB2 group. The motif GGACU was enriched significantly in both the control group and the CPB2 group. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analysis showed that the differentially methylated modified mRNAs were mainly enriched in Hippo signaling pathway and Wnt signaling pathway. In addition, the target genes of the differentially m6A modified lncRNAs were related to defense response to virus and immune response. For example, ENSSSCG00000042575, ENSSSCG00000048701 and ENSSSCG00000048785 might regulate the defense response to virus, immune and inflammatory response to resist the harmful effects of viruses on cells.
Conclusion:
In summary, this study established the m6A transcription profile of mRNAs and lncRNAs in IPEC-J2 cells treated by CPB2 toxin. Further analysis showed that m6A-modified RNAs were related to defense against viruses and immune response after CPB2 toxin treatment of the cells. Threem6A-modified lncRNAs, ENSSSCG00000042575, ENSSSCG00000048785 and ENSSSCG00000048701, were most likely to play a key role in CPB2 toxin-treated IPEC-J2 cells. The results provide a theoretical basis for further research on the role of m6A modification in piglet diarrhea.
Insights
This study reveals n6-methyladenosine (m6A) RNA modifications in piglet intestinal cells challenged with Clostridium perfringens type C toxin. Key m6A-modified lncRNAs are implicated in antiviral and immune responses, offering insights into infectious diarrhea.
Area of Science:
- Molecular Biology
- Epigenetics
- Animal Science
Background:
- n6-methyladenosine (m6A) modification is prevalent in RNA and linked to disease.
- The role of m6A in Clostridium perfringens type C infectious diarrhea is not well understood.
Purpose of the Study:
- To investigate the m6A methylation profiles of mRNAs and lncRNAs in an in vitro model of C. perfringens type C infectious diarrhea.
- To identify potential roles of m6A-modified RNAs in the cellular response to C. perfringens beta2 (CPB2) toxin.
Main Methods:
- Established an in vitro model using intestinal porcine jejunum epithelial cells (IPEC-J2) treated with CPB2 toxin.
- Employed methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) to analyze m6A modification and gene expression.
Main Results:
- Identified thousands of m6A-modified mRNAs and lncRNAs, with many showing differential expression upon CPB2 toxin treatment.
- Enrichment analysis revealed involvement in Hippo and Wnt signaling pathways for mRNAs and antiviral/immune responses for lncRNAs.
- Highlighted three specific m6A-modified lncRNAs (ENSSSCG00000042575, ENSSSCG00000048701, ENSSSCG00000048785) potentially crucial for cellular defense.
Conclusions:
- Established the m6A epitranscriptomic profile in IPEC-J2 cells responding to CPB2 toxin.
- m6A-modified RNAs are significantly associated with antiviral and immune responses in this model.
- Identified key lncRNAs that may play critical roles in mitigating CPB2 toxin effects, providing a basis for further research on piglet diarrhea.
