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Affinity Purification of the RNA Degradation Complex, the Exosome, from HEK-293 Cells.


ABSTRACT: The RNA exosome complex plays a central role in RNA processing and regulated turnover. Present both in cytoplasm and nucleus, the exosome functions through associations with ribonucleases and various adapter proteins (reviewed in [Kilchert et al., 2016]). The following protocol describes an approach to purify RNA exosome complexes from HEK-293 cells, making use of inducible ectopic expression, affinity capture, and rate-zonal centrifugation. The obtained RNA exosomes have been used successfully for proteomic, structural, and enzymatic studies (Domanski et al., 2016).

SUBMITTER: Domanski M 

PROVIDER: S-EPMC5499702 | biostudies-other | 2017 Apr

REPOSITORIES: biostudies-other

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