%0 Journal Article
%T A translational repression reporter assay for the analysis of RNA-binding protein consensus sites.
%A Nowacki J
%A Malenica M
%A Schmeing S
%A Schiller D
%A Buchmuller B
%A Amrahova G
%A 't Hart P
%J RNA Biol
%V 20
%N 1
%D 01 2023
%M 36946649
%F 4.766
%R 10.1080/15476286.2023.2192553
%X RNA-binding proteins are essential regulators of RNA processing and function. Translational repression assays can be used to study how they interact with specific RNA sequences by insertion of such a consensus sequence into the 5' untranslated region of a reporter mRNA and measuring reporter protein translation. The straightforward set-up of these translational repression assays avoids the need for the isolation of the protein or the RNA providing speed, robustness and a low-cost method. Here, we report the optimization of the assay to function with linear RNA sequences instead of the previously reported hairpin type sequences to allow the study of a wider variety of RNA-binding proteins. Multiplication of a consensus sequence strongly improves the signal allowing analysis by both fluorescence intensity measurements and flow cytometry.