Cas13b is a Type VI B CRISPR associated RNA Guided RNase differentially regulated by accessory proteins Csx27 and Csx28
CRISPR Cas adaptive immune systems defend microbes against foreign nucleic acids via RNA guided endonucleases. Using a computational sequence database mining approach, we identify two Class 2 CRISPR Cas systems (subtype VI B) that lack Cas1 and Cas2 and encompass a single large effector protein, Cas13b, along with one of two previously uncharacterized associated proteins, Csx27 or Csx28. We establish that these CRISPR Cas systems can achieve RNA interference when heterologously expressed. Through a combination of biochemical and genetic experiments, we show that Cas13b processes its own CRISPR array with short and long direct repeats, cleaves target RNA, and exhibits collateral RNase activity. Using an E. coli essential gene screen, we demonstrate that Cas13b has a double sided protospacer flanking sequence and elucidate RNA secondary structure requirements for targeting. We also find that Csx27 represses, whereas Csx28 enhances, Cas13b mediated RNA interference. Characterization of these CRISPR systems creates opportunities to develop tools to manipulate and monitor cellular transcripts.