7VTI

Crystal structure of the Cas13bt3-crRNA binary complex

Summary for 7VTI

• Entry DOI: 10.2210/pdb7vti/pdb
• Descriptor: Cas13bt3, crRNA, CHLORIDE ION, ... (7 entities in total)
• Functional Keywords: rna binding protein-rna complex, rna binding protein/rna
• Biological source: Planctomycetes bacterium; More
• Total number of polymer chains: 2
• Total formula weight: 105963.95

Authors

Nakagawa, R.,Takeda, N.S.,Tomita, A.,Hirano, H.,Kusakizako, T.,Nishizawa, T.,Yamashita, K.,Nishimasu, H.,Nureki, O. (deposition date: 2021-10-29, release date: 2022-08-24, Last modification date: 2024-11-13)

Primary citation

Nakagawa, R.,Kannan, S.,Altae-Tran, H.,Takeda, S.N.,Tomita, A.,Hirano, H.,Kusakizako, T.,Nishizawa, T.,Yamashita, K.,Zhang, F.,Nishimasu, H.,Nureki, O.
Structure and engineering of the minimal type VI CRISPR-Cas13bt3.
Mol.Cell, 82:3178-3192.e5, 2022

PubMed Abstract: Type VI CRISPR-Cas13 effector enzymes catalyze RNA-guided RNA cleavage and have been harnessed for various technologies, such as RNA detection, targeting, and editing. Recent studies identified Cas13bt3 (also known as Cas13X.1) as a miniature Cas13 enzyme, which can be used for knockdown and editing of target transcripts in mammalian cells. However, the action mechanism of the compact Cas13bt3 remains unknown. Here, we report the structures of the Cas13bt3-guide RNA complex and the Cas13bt3-guide RNA-target RNA complex. The structures revealed how Cas13bt3 recognizes the guide RNA and its target RNA and provided insights into the activation mechanism of Cas13bt3, which is distinct from those of the other Cas13a/d enzymes. Furthermore, we rationally engineered enhanced Cas13bt3 variants and ultracompact RNA base editors. Overall, this study improves our mechanistic understanding of the CRISPR-Cas13 enzymes and paves the way for the development of efficient Cas13-mediated transcriptome modulation technologies.

PubMed: 36027912
DOI: 10.1016/j.molcel.2022.08.001

Experimental method

X-RAY DIFFRACTION (1.89 Å)