Titlebook: Alternative Splicing; Methods and Protocol Peter Scheiffele,Oriane Mauger Book 2022 The Editor(s) (if applicable) and The Author(s), under

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Tethered Function Assays to Elucidate the Role of RNA-Binding Proteins, (MS2CP) with high affinity and specificity and by doing so, the POI is tethered to the reporter RNA. Here, we describe how with the help of this assay the human cytoplasmic poly(A) binding protein is recruited to a mini-μ RNA reporter, thereby influencing the stability of the reporter transcript.

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,An Optimized Protocol for the Mapping of Cell Type–Specific Ribosome-Associated Transcript Isoformsion and cellular functions. The isolation of ribosome-associated transcripts is a powerful approach for deep profiling of cell type–specific transcripts, and particularly well-suited for quantitative analysis of transcript isoforms. This method employs conditional ribosome epitope-tagging in genetic

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,FACS-Based Neuronal Cell Type–Specific RNA Isolation and Alternative Splicing Analysis,ributes to the marvelous complexity of the transcriptome in neurons. Given the differential expression of alternative splicing regulators and diversity in alternative splicing programs in neuronal subpopulations, it is urgent and necessary to develop methods to efficiently isolate diverse subgroups

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,Quantitative Measurement of Alternatively Spliced RNA Isoform Levels,ps) that include versus exclude a particular alternative RNA segment. The ratio measurement to study alternative splicing regulation can be confounded when alternative isoforms undergo differential RNA decay, for example, nonsense-mediated mRNA decay (NMD). Isoform-centric quantification is more inf

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Analysis of Splicing Regulation by Third-Generation Sequencing,cent transcripts. The regulation of exon inclusion by alternative splicing is one of the main sources of this diversity, which leads to the expansion of the proteome. The portfolio of alternative transcripts remains largely underestimated. Improvement of the sequencing technologies has enhanced the

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