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The ubiquitously expressed RNA-binding proteins Roquin-1 and Roquin-2 are essential for appropriate immune cell function and postnatal survival of mice.
A targeted si RNA screen to identify cofactors of Roquin-mediated post-transcriptional gene regulation.However, it is unknown if other cellular proteins contribute to target control.To identify cofactors of Roquin, we used RNA interference to screen ~1500 genes involved in RNA-binding or m RNA degradation, and identified NUFIP2 as a cofactor of Roquin-induced m RNA decay.1a, b), we observed strong downregulation of ICOS protein levels after doxycycline-induced Roquin-1 and m Cherry expression (Fig. si RNA-mediated depletion of Roquin resulted in derepression of ICOS (Fig. The assay was both robust and reproducible, as indicated by a Z′ factor of 0.7 (Fig.1d), which reflects the suitability of a given assay for high-throughput screening.