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  • Systematic Detection of Poly(A)+ RNA-Interacting Proteins and Their Differential Binding
    Milek, Miha, 1981- ; Landthaler, Markus
    RNA-binding proteins are dynamic posttranscriptional regulators of gene expression. Identification of mRNA-binding proteins in a given experimental setting is thus of great importance. We describe a ... procedure to enrich for direct poly(A)+ RNA protein binders by 4-thiouridine-enhanced UV cross-linking and oligo(dT) purification. Subsequent nuclease-mediated release of RNA-binding proteins (RBPs) from mRNA allows for detection of eluted proteins by mass spectrometry. In addition, we provide a comparative approach to detect differences in RBP binding activity upon a biological stimulus.
    Type of material - e-article
    Publish date - 2018
    Language - english
    COBISS.SI-ID - 33905369