Reconstitution of the Human Nuclear RNA Exosome

We describe procedures to clone, express, and reconstitute an active human nuclear RNA exosome. Individual recombinant subunits are expressed from E. coli and successfully reconstituted into the nuclear complex, which contains the noncatalytic nine-subunit exosome core, the endoribonuclease and exoribonuclease DIS3, the distributive exoribonuclease EXOSC10, the cofactors C1D and MPP6, and the RNA helicase MTR4.