6AI6
Crystal structure of SpCas9-NG
Summary for 6AI6
| Entry DOI | 10.2210/pdb6ai6/pdb |
| Descriptor | CRISPR-associated endonuclease Cas9/Csn1, RNA (81-MER), DNA (28-MER), ... (8 entities in total) |
| Functional Keywords | nuclease, hydrolase-rna-dna complex, hydrolase, hydrolase/rna/dna |
| Biological source | Streptococcus pyogenes serotype M1 More |
| Total number of polymer chains | 4 |
| Total formula weight | 197022.33 |
| Authors | Nishimasu, H.,Hirano, S.,Ishitani, R.,Nureki, O. (deposition date: 2018-08-21, release date: 2018-10-31, Last modification date: 2023-11-22) |
| Primary citation | Nishimasu, H.,Shi, X.,Ishiguro, S.,Gao, L.,Hirano, S.,Okazaki, S.,Noda, T.,Abudayyeh, O.O.,Gootenberg, J.S.,Mori, H.,Oura, S.,Holmes, B.,Tanaka, M.,Seki, M.,Hirano, H.,Aburatani, H.,Ishitani, R.,Ikawa, M.,Yachie, N.,Zhang, F.,Nureki, O. Engineered CRISPR-Cas9 nuclease with expanded targeting space Science, 361:1259-1262, 2018 Cited by PubMed Abstract: The RNA-guided endonuclease Cas9 cleaves its target DNA and is a powerful genome-editing tool. However, the widely used Cas9 enzyme (SpCas9) requires an NGG protospacer adjacent motif (PAM) for target recognition, thereby restricting the targetable genomic loci. Here, we report a rationally engineered SpCas9 variant (SpCas9-NG) that can recognize relaxed NG PAMs. The crystal structure revealed that the loss of the base-specific interaction with the third nucleobase is compensated by newly introduced non-base-specific interactions, thereby enabling the NG PAM recognition. We showed that SpCas9-NG induces indels at endogenous target sites bearing NG PAMs in human cells. Furthermore, we found that the fusion of SpCas9-NG and the activation-induced cytidine deaminase (AID) mediates the C-to-T conversion at target sites with NG PAMs in human cells. PubMed: 30166441DOI: 10.1126/science.aas9129 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.7 Å) |
Structure validation
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