Project description:The goal of this study is to investigate how WTAP regulates islet β-cell function. Islets were isolated from pancreatic islets of Wtapflox/flox and Wtap-βKO mice at 7 weeks old. One islet sample was combined from three mice. Total RNA was extracted using Tripure Isolation Reagent (Roche, Mannheim, Germany). Three independent biological replicates for each group were used for RNA-seq. RNA-seq was performed by deep sequencing using an Illumina Novaseq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome(GRCm38/mm10) with Hisat2 v2.0.5, and the aligned reads were used to quantify mRNA expression by using featureCounts v1.5.0-p3. Our study represents the first detailed analysis of islet transcriptomes from Wtapflox/flox and Wtap-βKO mice, generated by RNA-seq technology. The RNA-seq analysis showed that 3015 genes were downregulated and 2900 genes were upregulated in the pancreatic islets of Wtap-βKO mice.
Project description:Purpose: The goal of this study is to compare transcriptome profilings of interscapular brown adipose tissue (iBAT) from BAT-specific WTAP knockout and control mice Methods: iBATs were collected from WTAP BKO and f/f mice at 8 weeks old (n=3 for each group), and total RNA was isolated using TriPure. RNA-seq was performed by using Illumina NovaSeq 6000 platform. Paired-end clean reads were aligned to the mouse reference genome (Ensemble_grcm38_p6) with TopHat (version 2.0.12), and the aligned reads were used to quantify mRNA expression by using HTSeq-count (version 0.6.1). Conclusion:Our study represents the first detailed analysis of iBAT transcriptomes from WTAP BKO and f/f mice, generated by RNA-seq technology. The RNA-seq analysis showed that 894 genes were down-regulated and 1410 genes were up-regulated in the iBAT of WTAP BKO mice. GO analysis showed that the down-regulated genes were primarily related to related to generation of precursor metabolites and energy, cellular respiration, energy derivation by oxidation of organic compounds, electron transport chain, nucleotide metabolic process and purine ribonucleotide metabolic process.
Project description:Knock-down or overexpression of WTAP regulated migration and invasion of cholangiocarcinoma cells in vivo and vitro studies. To investigate the underlying mechanism for WTAP-regulated migration and invasion, the gene expression between the mock cells and the stable cells was compared Total RNA was purified from the mock cells and the stable cells overexpressing WTAP
Project description:Acute myeloid leukemia (AML) continues to have the lowest survival rates of all leukemias. Therefore, new therapeutic strategies are urgently needed to improve clinical outcomes for AML patients. Here, we report a novel role for Wilms’ tumor 1-associated protein (WTAP) in pathogenesis of AML. We have performed RNA-Seq in K562 cells with knockdown of WTAP to ascertain which genes it regulates. We have 2 replicates of total RNA for K562 cells and 2 replicates with WTAP knocked down
Project description:The goal of this project is to investigate the role of SIRT1, the most conserved mammalian NAD+-dependent protein deacetylase, in the regulation of heart development. SIRT1 is important for heart development and functions. However, the underlying molecular mechanisms remain undefined. In this study, we analyzed the gene expression profiles in E18.5 WT and SIRT1 KO mouse hearts.
Project description:label the cells overexpressed Myc tagged METTL3 and Flag tagged WTAP with 4-SU, the RNA bound by METT3,WTAP can be got by Myc or Flag IP followed by RNA isolation by using the TRIzol (Invitrogen) reagent by following the company manual.the RNA was then used for library preparation using a TruSeq™ RNA Sample Prep Kit v2 (Illumina, San Diego, CA, USA) according to the manufacturer’s protocol.The libraries were sequenced using HiSeq2000 (Illumina) in single-read mode, creating reads with a length of 50 bp. Sequencing chemistry v2 (Illumina) was used and samples were multiplexed in two samples per lane. discovery of the binding motif of METTL3，WTAP in METTL3，WTAP overexpressed Human 293T cells
