9IZQ
Cryo-EM structure of CasLambda2-crRNA-target DNA ternary complex in the intermediate state
Summary for 9IZQ
| Entry DOI | 10.2210/pdb9izq/pdb |
| EMDB information | 61039 |
| Descriptor | Cas Lambda2, RNA (58-MER), DNA (40-MER), ... (5 entities in total) |
| Functional Keywords | crispr-cas12, genome engineering, hydrolase-rna-dna complex, dna binding protein |
| Biological source | unidentified More |
| Total number of polymer chains | 4 |
| Total formula weight | 132497.53 |
| Authors | Omura, S.N.,Hirano, H.,Itoh, Y.,Nureki, O. (deposition date: 2024-08-01, release date: 2025-06-11, Last modification date: 2025-06-25) |
| Primary citation | Omura, S.N.,Alfonse, L.E.,Ornstein, A.,Morinaga, H.,Hirano, H.,Itoh, Y.,Munoz, G.,Garrity, A.J.,Hoffman, G.R.,DiTommaso, T.,Yan, W.X.,Cheng, D.R.,Scott, D.A.,Maben, Z.,Nureki, O. Structural basis for target DNA cleavage and guide RNA processing by CRISPR-Cas lambda 2. Commun Biol, 8:876-876, 2025 Cited by PubMed Abstract: RNA-guided CRISPR-Cas nucleases are widely used as versatile genome-engineering tools. Among the diverse CRISPR-Cas effectors, CRISPR-Casλ-also referred to as Cas12n-is a recently identified miniature type V nuclease encoded in phage genomes. Given its demonstrated nuclease activity in both mammalian and plant cells, Casλ has emerged as a promising candidate for genome-editing applications. However, the precise molecular mechanisms of Casλ family enzymes remain poorly understood. In this study, we report the identification and detailed biochemical and structural characterizations of CRISPR-Casλ2. The cryo-electron microscopy structures of Casλ2 in five different functional states unveiled the dynamic domain rearrangements during its activation. Our biochemical analyses indicated that Casλ2 processes its precursor crRNA to a mature crRNA using the RuvC active site through a unique ruler mechanism, in which Casλ2 defines the spacer length of the mature crRNA. Furthermore, structural comparisons of Casλ2 with Casλ1 and CasΦ highlighted the diversity and conservation of phage-encoded type V CRISPR-Cas enzymes. Collectively, our findings augment the mechanistic understanding of diverse CRISPR-Cas nucleases and establish a framework for rational engineering of the CRISPR-Casλ-based genome-editing platform. PubMed: 40473912DOI: 10.1038/s42003-025-08300-8 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.06 Å) |
Structure validation
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