Summary for 7TR9
| Entry DOI | 10.2210/pdb7tr9/pdb |
| EMDB information | 26083 |
| Descriptor | Cas8a, NICKEL (II) ION, Cas11a, ... (10 entities in total) |
| Functional Keywords | crispr, cascade, cas3, type i-a, genome editing, rna binding protein |
| Biological source | Pyrococcus furiosus DSM 3638 More |
| Total number of polymer chains | 19 |
| Total formula weight | 499218.79 |
| Authors | Hu, C.,Ni, D.,Nam, K.H.,Majumdar, S.,McLean, J.,Stahlberg, H.,Terns, M.,Ke, A. (deposition date: 2022-01-28, release date: 2022-08-10, Last modification date: 2024-10-16) |
| Primary citation | Hu, C.,Ni, D.,Nam, K.H.,Majumdar, S.,McLean, J.,Stahlberg, H.,Terns, M.P.,Ke, A. Allosteric control of type I-A CRISPR-Cas3 complexes and establishment as effective nucleic acid detection and human genome editing tools. Mol.Cell, 82:2754-2768.e5, 2022 Cited by PubMed Abstract: Type I CRISPR-Cas systems typically rely on a two-step process to degrade DNA. First, an RNA-guided complex named Cascade identifies the complementary DNA target. The helicase-nuclease fusion enzyme Cas3 is then recruited in trans for processive DNA degradation. Contrary to this model, here, we show that type I-A Cascade and Cas3 function as an integral effector complex. We provide four cryoelectron microscopy (cryo-EM) snapshots of the Pyrococcus furiosus (Pfu) type I-A effector complex in different stages of DNA recognition and degradation. The HD nuclease of Cas3 is autoinhibited inside the effector complex. It is only allosterically activated upon full R-loop formation, when the entire targeted region has been validated by the RNA guide. The mechanistic insights inspired us to convert Pfu Cascade-Cas3 into a high-sensitivity, low-background, and temperature-activated nucleic acid detection tool. Moreover, Pfu CRISPR-Cas3 shows robust bi-directional deletion-editing activity in human cells, which could find usage in allele-specific inactivation of disease-causing mutations. PubMed: 35835111DOI: 10.1016/j.molcel.2022.06.007 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.9 Å) |
Structure validation
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