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RNA whole-mount In Situ Hybridization Proximity Ligation Assay (rISH-PLA), an assay for detecting RNA-protein complexes in intact cells

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

Techniques for studying RNA-protein interactions have lagged behind those for DNA-protein interactions as a consequence of the complexities associated with working with RNA. This unit describes a method for the adaptation of the In Situ Hybridization-Proximity Ligation Assay (ISH-PLA) to the study of RNA regulation (rISH-PLA). The rISH-PLA assay allows the identification of a given RNA-protein complex at subcellular and single-cell resolution, thus avoiding the lack of spatial resolution and sensitivity associated with assaying heterogeneous cell populations from which conventional RNA-protein interaction detection techniques suffer. This technique will be particularly usefully for studying the activity of RNA binding proteins (RBPs) in complex mixtures of cells, for example tissue sections or whole embryos.

Original languageEnglish
Title of host publicationCurrent Protocols in Cell Biology
EditorsAditi Nadkami
PublisherWiley Online Library
Volume74
ISBN (Electronic)978-0471143031
DOIs
Publication statusEarly online - 3 Mar 2017

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