8WF9

Cryo-EM structure of the PspCas13b-crRNA-target RNA complex (State 1)

Summary for 8WF9

• Entry DOI: 10.2210/pdb8wf9/pdb
• EMDB information: 37487
• Descriptor: PspCas13b, crRNA, target RNA, ... (4 entities in total)
• Functional Keywords: crispr, rnase, rna binding protein-rna complex, rna binding protein/rna
• Biological source: Prevotella sp.; More
• Total number of polymer chains: 3
• Total formula weight: 160803.02

Authors

Ishikawa, J.,Kato, K.,Yamashita, K.,Nishizawa, T.,Nishimasu, H. (deposition date: 2023-09-19, release date: 2025-03-19, Last modification date: 2025-04-23)

Primary citation

Ishikawa, J.,Kato, K.,Kannan, S.,Okazaki, S.,Ishiguro, S.,Yamashita, K.,Yachie, N.,Nishizawa, T.,Zhang, F.,Nishimasu, H.
Structural insights into RNA-guided RNA editing by the Cas13b-ADAR2 complex.
Nat.Struct.Mol.Biol., 2025

PubMed Abstract: Cas13 is an RNA-guided RNA endonuclease derived from the type VI CRISPR-Cas system, which has been used in numerous RNA-targeting technologies, such as RNA knockdown, detection and editing. The catalytically inactive Prevotella sp. Cas13b (dPspCas13b) fused to the human adenosine deaminase acting on RNA 2 (ADAR2) deaminase domain can edit adenosine in target transcripts to inosine, in an RNA-editing technology called REPAIR (RNA editing for programmable A-to-I replacement), which has potential for gene therapy. Here we report the cryo-electron microscopy structures of the PspCas13b-guide RNA binary complex, the PspCas13b-guide RNA-target RNA ternary complex and the dPspCas13b-ADAR2-guide RNA-target RNA complex. These structures provide mechanistic insights into RNA cleavage and editing. We applied our structural insights to engineer a compact and efficient dPspCas13b-ADAR2 complex (REPAIR-mini). Overall, our findings advance the understanding of CRISPR-Cas13 effector nucleases and could enable the development of improved RNA-targeting technologies.

PubMed: 40217120
DOI: 10.1038/s41594-025-01529-1

Experimental method

ELECTRON MICROSCOPY (3.13 Å)