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Global Identification of Human Exosome Substrates Using RNA Interference and RNA Sequencing

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DOI

The RNA exosome is involved in RNA processing and quality control. In humans, it consists of an enzymatically inactive nine-subunit core, with ribonucleolytic activity contributed by one or two additional components. Moreover, several protein cofactors interact with the exosome to enable and specify its recruitment to a wide range of substrates. A common strategy to identify these substrates has been to deplete an exosome subunit or a cofactor and subsequently interrogate which transcripts become stabilized. Here, we describe an experimental pipeline including siRNA-mediated depletion of the RNA exosome or its cofactors in HeLa cells, confirmation of the knockdown efficiencies, and the manual or high-throughput identification of exosome targets.
OriginalsprogEngelsk
TitelThe Eukaryotic RNA Exosome : Methods and Protocols
Antal sider19
ForlagHumana Press
Udgivelsesår2020
Sider127-145
DOI
StatusUdgivet - 2020
SerietitelMethods in Molecular Biology
Vol/bind2062
ISSN1064-3745

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