Project description:Alkbh5/Mettl3loxp/loxp mice were crossed with Lyz2-Cre mice to generate macrophage-specific conditional knockout mice for Alkbh5/Mettl3. Bone marrow-derived macrophages (BMDMs) were isolated from these mice, and MeRIP-seq was performed to investigate the impact of the knockout on RNA N6-Methyladenosine
Project description:To identify potential target mRNAs that are demethylated by ALKBH5, meRIP-seq with a m6A antibody was conducted in 143B osteosarcoma cells transfected with scrambled or ALKBH5-siRNA.
Project description:We performed MeRIP-seq of METTL3-knockdown (KD) MIAPaCa-2 and control cells to detect METTL3-methylated genes and extracted genes whose methylation was lost in METTL3-KD cells compared with control cells. Next, we performed mRNA expression analysis using dates of input samples at MeRIP-seq. Genes whose methylation peaks were abolished by METTL3-KD and whose expression changes were observed were analyzed as candidates for METTL3-regulated genes.
Project description:To determine the targets underlying ALKBH5 during head and neck squamouse cell carcinoma progression, Methylated RNA immunoprecipitation (MeRIP) with an m6A specific antibody followed by RNA sequencing (MeRIP-seq) and next generation sequencing were combined to screen the potential targets haboring m6A modificatios and mRNA level alteration after ALKBH5 knockdown in a HNSCC cell line.
Project description:By performing RNA-seq analysis on bone marrow neutrophils from the Alkbh5-deficient mice and Wild-type littermates undergoing CLP-induced sepsis, we want to investigate the effect of ALKBH5 on transcriptional landscape of mouse bone marrow neutrophils during bacterial infection. Then, we performed gene expression profiling and Gene Ontology enrichment analysis of the significantly differentially expressed genes using data obtained from RNA-seq.
Project description:To identify the molecular mechanism by which METTL3 regulates endothelial barrier function, we performed RNA-seq and MeRIP-seq in HULEC-5a cells with stable METTL3 knockdown and control cells. The RNA-seq results revealed that 437 transcripts were significantly downregulated (fold change <0.5) after METTL3 knockdown. The MeRIP-seq results revealed that the m6A peaks in 1011 transcripts were decreased in abundance (fold change >1.2). Intriguingly, 55 transcripts overlapped in the RNA-seq and MeRIP-seq data
Project description:We performed MeRIP-seq on (1) two human epidermis tissue and HaCAT (human keratinocyte cell line) cells; (2) HaCAT cell lines with or without ALKBH5 knockdown by siRNA in duplicate experiments.
Project description:Mettl3 is an mRNA methyl transferase enzyme. It catalzyes the methylation of the most common mRNA biochemical modification N6-methyl adenosine (m6A). The role of Mettl3/m6A pathway is unknown in the developing kidney. Through genetic and pharmaceutical approaches we found that Mettl3 is required for the differentiation of nephron progenitor cells. nephron progenitors. Genetic deletion of Mettl3 in the nephron progenitor cells blocks their differentiation. We sought to identify the mechanism of Mettl3 activity. We performed RNA-sequencing and RNA-m6A pull down assay followed by next-generation sequencing (MeRIP-seq) on wild type and Mettl3 deleted kidneys to determine the mRNA targets of Mettl3.
Project description:Hypertrophic scars (HTS) are fibrotic skin disorders characterized by excessive deposition of dermal extracellular matrix (ECM), presenting significant challenges in clinical management despite incomplete understanding of their underlying mechanisms. Using MeRIP-seq and RNA-seq, our study initially revealed the direct regulatory role of ALKBH5 on ECM components in dermal fibroblasts, thereby involving in the pathogenesis of HTS.