关键词: 3′ Untranslated regions (UTRs) Individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP) Intron Primary B cells RNA binding proteins mRNA maturation mRNA stability mRNA translation

Mesh : Animals B-Lymphocytes / immunology metabolism Binding Sites Cell Separation / methods Computational Biology / methods Gene Expression Profiling / methods Gene Library High-Throughput Nucleotide Sequencing Humans Immunoprecipitation / methods Introns Lymphocyte Activation Protein Binding RNA Stability RNA-Binding Proteins / metabolism Transcriptome Ultraviolet Rays

来  源:   DOI:10.1007/978-1-4939-7095-7_14

Abstract:
Posttranscriptional regulation of gene expression shapes the B cell transcriptome and controls messenger RNA (mRNA) translation into protein. Recent reports have highlighted the importance of RNA binding proteins (RBPs) for mRNA splicing, subcellular location, stability, and translation during B lymphocyte development, activation, and differentiation. Here we describe individual-nucleotide resolution UV cross-linking and immunoprecipitation (iCLIP) in primary lymphocytes, a method that maps RNA-protein interactions in a genome-wide scale allowing mechanistic analysis of RBP function. We discuss the latest improvements in iCLIP technology and provide some examples of how integration of the RNA-protein interactome with other high-throughput mRNA sequencing methodologies uncovers the important role of RBP-mediated RNA regulation in key biological cell processes.
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