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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Quantitative Detection of Protein Splice Variants by Selected Reaction Monitoring (SRM) Mass Spectr tools to specifically detect and quantify proteoforms (Smith et al., Nat Methods 10:186–187, 2013) is a major impediment to functional studies. Recently, biological mass spectrometry (MS) has undergone impressive advances (Mann, Nat Rev Mol Cell Biol 17:678, 2016), including the generation of a hig

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Genome-Wide CRISPR Screening to Identify Mammalian Factors that Regulate Intron Retention,a wide variety of genes controlled by IR, few techniques are available to identify regulators of IR in an unbiased manner. Here, we describe a CRISPR knockout screening method that can be applied to uncover regulators of IR. This method uses GFP reporter constructs containing a retained intron from

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Tethered Function Assays to Elucidate the Role of RNA-Binding Proteins, To elucidate the effect of a specific RBP on bound RNA, it can be artificially recruited to a specific site on a reporter mRNA that can be followed by a variety of methods. In this so-called tethering assay, the protein of interest (POI) is fused to the coat protein of the MS2 bacteriophage and exp