Structure of a type IV CRISPR-Cas ribonucleoprotein complex

We reveal the cryo-electron microscopy structure of a type IV-B CRISPR ribonucleoprotein (RNP) complex (Csf) at 3.9-Å resolution. The complex best resembles the type III-A CRISPR Csm effector complex, consisting of a Cas7-like (Csf2) filament intertwined with a small subunit (Cas11) filament, but the complex lacks subunits for RNA processing and target DNA cleavage. Surprisingly, instead of assembling around a CRISPR-derived RNA (crRNA), the complex assembles upon heterogeneous RNA of a regular length arranged in a pseudo-A-form configuration. These findings provide a high-resolution glimpse into the assembly and function of enigmatic type IV CRISPR systems, expanding our understanding of class I CRISPR-Cas system architecture, and suggesting a function for type IV-B RNPs that may be distinct from other class 1 CRISPR-associated systems. [Display omitted] •The type IV-B CRISPR-Cas Csf proteins assemble around an RNA•First structure of a type IV-B CRISPR-Cas RNP complex•Structure confirms evolutionary link of type IV systems from a type III-like ancestor•Assembly of RNP complex on non-CRISPR RNAs suggests a non-canonical role Biological Sciences; Structural Biology