{Reference Type}: Journal Article
{Title}: A translational repression reporter assay for the analysis of RNA-binding protein consensus sites.
{Author}: Nowacki J;Malenica M;Schmeing S;Schiller D;Buchmuller B;Amrahova G;'t Hart P;
{Journal}: RNA Biol
{Volume}: 20
{Issue}: 1
{Year}: 01 2023
{Factor}: 4.766
{DOI}: 10.1080/15476286.2023.2192553
{Abstract}: 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.