6DTD

High-resolution crystal structure of Cas13b from Prevotella buccae

Summary for 6DTD

• Entry DOI: 10.2210/pdb6dtd/pdb
• Descriptor: nuclease, RNA (37-MER), CITRIC ACID, ... (7 entities in total)
• Functional Keywords: nuclease, hydrolase-rna complex, hydrolase/rna
• Biological source: Prevotella buccae; More
• Total number of polymer chains: 2
• Total formula weight: 148288.56

Authors

Slaymaker, I.M.S.,Zhang, F.Z. (deposition date: 2018-06-15, release date: 2019-02-20, Last modification date: 2024-03-13)

Primary citation

Slaymaker, I.M.,Mesa, P.,Kellner, M.J.,Kannan, S.,Brignole, E.,Koob, J.,Feliciano, P.R.,Stella, S.,Abudayyeh, O.O.,Gootenberg, J.S.,Strecker, J.,Montoya, G.,Zhang, F.
High-Resolution Structure of Cas13b and Biochemical Characterization of RNA Targeting and Cleavage.
Cell Rep, 26:3741-3751.e5, 2019

PubMed Abstract: Type VI CRISPR-Cas systems contain programmable single-effector RNA-guided RNases, including Cas13b, one of the four known family members. Cas13b, which has been used for both RNA editing and nucleic acid detection, is unique among type VI CRISPR effectors in its linear domain architecture and CRISPR RNA (crRNA) structure. Here, we report the crystal structure of Prevotella buccae Cas13b (PbuCas13b) bound to crRNA at 1.65 Å resolution. This structure, combined with biochemical experiments assaying the stability, kinetics, and function of Cas13b, provides a mechanistic model for Cas13b target RNA recognition and identifies features responsible for target and cleavage specificity. Based on these observations, we generated Cas13b variants with altered cleavage preferences, which may expand the utility of nuclease-based RNA detection assays and other applications of Cas13b in mammalian cells.

PubMed: 30917325
DOI: 10.1016/j.celrep.2019.02.094

Experimental method

X-RAY DIFFRACTION (1.65 Å)