8HUD
Cryo-EM structure of the EvCas9-sgRNA-target DNA ternary complex
Summary for 8HUD
| Entry DOI | 10.2210/pdb8hud/pdb |
| EMDB information | 35035 |
| Descriptor | CRISPR-associated endonuclease Cas9, sgRNA, Target DNA strand, ... (4 entities in total) |
| Functional Keywords | rna, dna, rna binding protein/rna/dna, rna binding protein-rna-dna complex |
| Biological source | Eubacterium ventriosum ATCC 27560 More |
| Total number of polymer chains | 4 |
| Total formula weight | 166195.96 |
| Authors | |
| Primary citation | Tang, N.,Wu, Z.,Gao, Y.,Chen, W.,Wang, Z.,Su, M.,Ji, W.,Ji, Q. Molecular Basis and Genome Editing Applications of a Compact Eubacterium ventriosum CRISPR-Cas9 System. Acs Synth Biol, 13:269-281, 2024 Cited by PubMed Abstract: CRISPR-Cas9 systems have been widely harnessed for diverse genome editing applications because of their ease of use and high efficiency. However, the large molecular sizes and strict PAM requirements of commonly used CRISPR-Cas9 systems restrict their broad applications in therapeutics. Here, we report the molecular basis and genome editing applications of a novel compact type II-A CRISPR-Cas9 system (EvCas9) with 1107 residues and distinct 5'-NNGDGN-3' (where D represents A, T, or G) PAM specificity. We determine the cryo-EM structure of EvCas9 in a complex with an sgRNA and a target DNA, revealing the detailed PAM recognition and sgRNA and target DNA association mechanisms. Additionally, we demonstrate the robust genome editing capacity of EvCas9 in bacteria and human cells with superior fidelity compared to SaCas9 and SpCas9, and we engineer it to be efficient base editors by fusing a cytidine or adenosine deaminase. Collectively, our results facilitate further understanding of CRISPR-Cas9 working mechanisms and expand the compact CRISPR-Cas9 toolbox. PubMed: 38061052DOI: 10.1021/acssynbio.3c00501 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.43 Å) |
Structure validation
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