Similar Datasets
Project description:IP with m6A-specific antibodies and input sequencing for every sample.
2021-04-06 | GSE171472 | GEO
Project description:Purpose: Evaluation of the m6A modification of PRV and PK15 transcripts during PRV infection Methods: Porcine kidney cell line PK15 was uninfected or infected with PRV for 24 hours. Total RNA from each sample were extracted. Intact mRNA was isolated from total RNA samples and then chemically fragmented to 300-nucleoside-long fragments. Fragmented mRNAs were immunoprecipitated with anti-N6-methyadenosine (m6A) antibody (a part of the fragmented mRNAs was kept as input). Both m6A enriched mRNAs and input mRNAs were concentrated for RNA-seq libraries construction. The libraries were forwarded to sequencing run on Illumina NovaSeq 6000. Results: PRV transcripts were m6A modified during PRV infection and PRV infection changed m6A modification profiles of PK15 transcripts.
2022-11-02 | GSE209949 | GEO
Project description:Purpose: Evaluation of the m6A modification of EBV and BJAB transcripts during EBV infection Methods: Human B lymphoma cell line BJAB was uninfected or infected with EBV for 24 hours. Total RNA from each sample were extracted. Intact mRNA was isolated from total RNA samples and then chemically fragmented to 100-nucleoside-long fragments. m6A methylated mRNAs were immunoprecipitated with anti-N6-methyadenosine (m6A) antibody (a part of the fragmented mRNAs was kept as input). Both m6A enriched mRNAs and input mRNAs were concentrated for RNA-seq library construction. Sequencing was performed using an Illumina HiSeq 4000. Results: EBV EBNA2 and BHRF1 transcripts were m6A modified and m6A modification of BJAB transcripts changed during EBV infection. Conclusions: Our study found that some EBV transcripts were m6A modified during EBV infection and EBV infection changed m6A modification profiles of BJAB transcripts.
2021-07-10 | GSE133936 | GEO
Project description:In this experiment, m6A-seq sequencing technology was used to study the functional role of methylated molecules in the process of CPB2 processing porcine small intestinal epithelial cells. In this experiment, an IP library and an input library were built together. The IP library was enriched with m6A specific antibodies to generate methylated RNA, and the influence of m6A methylation modification on its expression was analyzed by bioinformatics. We finally concluded that m6A methylation modification may play a very important role after CPB2 toxin treats small intestinal epithelial cells.
2021-10-05 | GSE167267 | GEO
Project description:In this study, we aimed to systematically profile global RNA N6-methyladenosine (m6A) modification patterns in a mouse model of diabetic cardiomyopathy (DCM). Patterns of m6A in DCM and normal hearts were analyzed via m6A-specific methylated RNA immunoprecipitation followed by high-throughput sequencing (MeRIP-seq) and RNA sequencing (RNA-seq). A total of 973 m6A peaks were detected in DCM samples and 296 differentially methylated sites were selected for further study, including 106 hypermethylated and 190 hypomethylated m6A sites (fold change (FC) > 2, p < 0.05). Gene ontology and KEGG Pathway analyses indicated that unique m6A-modified transcripts in DCM were closely linked to cardiac fibrosis, myocardial hypertrophy, and myocardial energy metabolism. Overall, m6A modification patterns were altered in DCM, and modification of epitranscriptomic processes, such as m6A, is a potentially interesting therapeutic approach.
2021-05-25 | GSE173384 | GEO
Project description:We report the application of MeRIP sequencing technology for high-throughput profiling of RNA m6A modifications in wide-type and knock-down METTL3 or WTAP Human Umbilical Vein Endothelial Cells (HUVECs). Both the input samples without immunoprecipitation and the m6A IP samples were used for RNA-seq library generation with NEBNext® Ultra II Directional RNA Library Prep Kit (New England Biolabs, Inc., USA). Library sequencing was performed on an illumina Hiseq instrument with 150bp paired-end reads. Clean reads of all libraries were aligned to the reference genome (HG19) by Hisat2 software (v2.0.4). Methylated sites on RNAs (peaks) were identified by MACS software. Differentially methylated sites were identified by diffReps. And, guided by the Ensembl gtf gene annotation file, cuffdiff software (part of cufflinks) was used to get the gene level FPKM as the expression profiles of mRNA, and fold change and p-value were calculated based on FPKM, differentially expressed mRNA were identified. qRT-PCR validation was performed using SYBR Green assays. Finally, we find that METTL3/WTAP can regulate the expression level of target genes through m6A modification in HUVECs. This study provides a framework for applying MeRIP sequencing profiles to characterize vascular endothelial cells.
2019-12-20 | GSE142386 | GEO
Project description:N6-methyladenosine (m6A) is the most prevalent modification in eukaryotic mRNA and potential regulatory functions of m6A have been shown by mapping the RNA m6A modification landscape. M6A modification in active gene regulation manifests itself as altered methylation profiles. However, the profiling of m6A modification and its potential role in gestational diabetes mellitus (GDM) has not yet been studied. In this work, placental samples were collected from GDM and control patients. MeRIP-seq was performed to identify differences in m6A methylation. Fragmented mRNAs were incubated for 2 h at 4 °C in the presence of 2 μg m6A antibodies (Synaptic Systems, 202003) in a 500 μl IP reaction system, and some of the fragments were used as input. RNA-seq libraries for m6A antibody-enriched mRNAs and input mRNAs were prepared using the KAPA Stranded mRNA-seq Kit (Illumina, CA, USA). Altered peaks of m6A-modified transcripts were primarily associated with mTOR signaling pathway, Notch signaling pathway, TGF-beta signaling pathway and so on. Our data provide novel information regarding m6A modification alterations in GDM and help our understanding of the pathogenesis of GDM.
2021-12-31 | GSE154413 | GEO
Project description:MeRIP-Seq data aligned to the genome (GRCh38) for cells with IDH1-Mut or IDH1-WT genotypes. Aligned data (BAM) are separated into input RNA and m6A immunoprecipitated RNA for each cell sample.
2024-02-19 | E-MTAB-13765 | biostudies-arrayexpress
Project description:Background: Abdominal aortic aneurysm (AAA) is a vascular disease with indeterminate prevalence but high mortality rates when complicated with rupture. The pathogenesis of AAA has not been fully elucidated. N6-methyladenosine (m6A) modification is likely important in the development of AAA. In the present study, m6A-MeRIP sequencing and RNA sequencing were performed to identify the m6A sites. Bioinformatics analysis was used to evaluate the m6A patterns of the aorta walls of AngII-induced abdominal aortic aneurysm (AAA) model and normal mice. Results: There were 2039 differentially methylated m6A peaks involving 1865 mRNAs in the AAA group relative to the control, of which 1610 peaks in 1466 mRNAs were hypermethylated and 429 peaks in 410 mRNAs were hypomethylated. The hypermethylated mRNAs in AAA group were mostly enriched in transcription regulation and intercellular signaling, especially the Wnt signaling-associated processes. Hypomethylated m6A sites were mainly enriched in G protein-coupled receptor activity and ion channel activity. Conclusion: Our study suggested an original viewpoint that AAA might mainly be relevant to combined effect of m6A methylation modification in Wnt pathway, G protein-coupled receptor and ion channel- associated genes, which were worthy of further investigation.
2021-04-09 | GSE171371 | GEO