biostudies-arrayexpressMetabolomicsUnknownTranscriptomicsGenomicsProteomicsThe regulation of alternative splicing involves interactions between RNA-binding proteins and pre-mRNA positions close to the splice sites. T-cell intracellular antigen 1 (TIA1) and TIA1-like 1 (TIAL1) locally enhance exon inclusion by recruiting U1 snRNP to 5' splice sites. However, effects of TIA proteins on splicing of distal exons have not yet been explored. We used UV-crosslinking and immunoprecipitation (iCLIP) to find that TIA1 and TIAL1 bind at the same positions on human RNAs. Binding downstream of 5' splice sites was used to predict the effects of TIA proteins in enhancing inclusion of proximal exons and silencing inclusion of distal exons. The predictions were validated in an unbiased manner using splice-junction microarrays, RT-PCR, and minigene constructs, which showed that TIRNAi profiling by arrayHomo sapiensHomo sapienshttps://www.ebi.ac.uk/biostudies/studies/E-MTAB-526biostudies-arrayexpressiCLIP Predicts the Dual Splicing Effects of TIA-RNA InteractionsSample Processing - HeLa cells are transfected with 0.5 nM siRNA oligo for TIA1 and TIAL1 gene each. 3 different pairs of oligos are used together with a scrambled control. Cells are transfected using RNAiMax at d1, and the media is changed to DMEM with 10% FBS without antibotics. The cells are re-transfected again at d3. The cells are collected at d5.Nucleic Acid Extraction - mRNAs are extracted using RNeasy kit from Qiagen.Labeling - The cDNAs for hybridisaion are labelled using Affymetrix terminal labelling kit.Growth Protocol - HeLa cells are grown in DMEM with 10% FBS and Penicillin/StreptomycinMIAME ScoreRaw DataAssays and DataAdditional FilesMAGE-TAB FilesArray DesignsZhen Wang, Melis Kayikci, Michael Briese, Kathi Zarnack, Nicholas M. Luscombe, Gregor Rot, Blaz Zupan, Tomaz Curk, Jernej Ulefalsemonitoring splicing changes in TIA1/TIAL1 double knockdown vs control HeLa cells2010-10-26T00:00:00Z2022-11-22T14:19:28.133Z2022-03-07T00:30:55.513ZE-MTAB-52621048981EFO_0001030