Comprehensive analysis of interacting RNAs with double-stranded RNA binding protein, TRBP. Homo sapiens

The goal of our project is to comprehensively identify the interacting long and small RNAs with double-stranded RNA binding protein, TRBP, to reveal the function of TRBP protein in RNA-associated phenomenon in human cells. We established a Tet-On inducible cell line that express N-terminal Flag-tagged wild-type (wt) and dsRBD3-deleted (delta_dsRBD3) TRBP, respectively, by the addition of doxycycline. TRBP is known to interact with Dicer through the dsRBD3, therefore delta_dsRBD3 TRBP mutant is assumed to lose the ability to interact with Dicer. We immunoprecipitated Flag-wt TRBP and Flag-delta_dsRBD3 TRBP using anti-FLAG antibody, and confirmed that wt TRBP interacts with Dicer but delta_dsRBD3 TRBP does not. Using such immunoprecipitated samples, we intend to identify the interacting long and small RNAs with wild-type and dsRBD3-deleted TRBP proteins, respectively.