Tethered Function Assays to Elucidate the Role of RNA-Binding Proteins.

Eberle, Andrea B; Mühlemann, Oliver (2022). Tethered Function Assays to Elucidate the Role of RNA-Binding Proteins. Methods in molecular biology, 2537, pp. 285-306. Springer 10.1007/978-1-0716-2521-7_17

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The fate of each RNA molecule is strongly determined by RNA-binding proteins (RBPs) which accompany transcripts from its synthesis to its degradation. To elucidate the effect of a specific RBP on bound RNA, it can be artificially recruited to a specific site on a reporter mRNA that can be followed by a variety of methods. In this so-called tethering assay, the protein of interest (POI) is fused to the coat protein of the MS2 bacteriophage and expressed in your favorite cells together with a reporter gene containing MS2 binding sites. The MS2 binding sites are recognized by the MS2 coat protein (MS2CP) with high affinity and specificity and by doing so, the POI is tethered to the reporter RNA. Here, we describe how with the help of this assay the human cytoplasmic poly(A) binding protein is recruited to a mini-μ RNA reporter, thereby influencing the stability of the reporter transcript.

Item Type:	Journal Article (Original Article)
Division/Institute:	08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP)
UniBE Contributor:	Eberle, Andrea Brigitte, Mühlemann, Oliver
Subjects:	500 Science > 570 Life sciences; biology 500 Science > 540 Chemistry
ISSN:	1940-6029
Publisher:	Springer
Language:	English
Submitter:	Pubmed Import
Date Deposited:	29 Jul 2022 07:29
Last Modified:	05 Dec 2022 16:22
Publisher DOI:	10.1007/978-1-0716-2521-7_17
PubMed ID:	35895271
Uncontrolled Keywords:	MS2 binding sites MS2 coat protein (MS2CP) Nonsense-mediated mRNA decay (NMD) Poly(A) binding protein RNA-binding protein Tethered protein Tethering assay
URI:	https://boris.unibe.ch/id/eprint/171588