UPF1 RNA Immunoprecipitation from Mini-μ Construct–expressing Cells

Zünd, David; Mühlemann, Oliver (2014). UPF1 RNA Immunoprecipitation from Mini-μ Construct–expressing Cells. Bio-protocol bio-protocol

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UPF1, an RNA helicase and a core factor of nonsense-mediated mRNA decay (NMD), interacts with RNA independently of the sequence context. To investigate the influence of translation on the association of UPF1 with specific reporter transcripts, UPF1 RNA immunoprecipitations (RIPs) are performed from Hela cells that either express a normally translated immunoglobulin-µ (Ig-µ) reporter (mini µ) or a version with a stable stem loop in the 5' UTR (SL mini µ) that efficiently inhibit translation initiation (Zund et al., 2013). Both the cloning of the SL mini µ reporter construct and the UPF1 RIP experiment are described in detail.

Item Type:

Journal Article (Original Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Zünd, David and Mühlemann, Oliver

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

2331-8325

Publisher:

bio-protocol

Language:

English

Submitter:

Christina Schüpbach

Date Deposited:

25 Jun 2014 09:54

Last Modified:

06 Dec 2014 03:11

BORIS DOI:

10.7892/boris.52145

URI:

https://boris.unibe.ch/id/eprint/52145

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