5B2P
Crystal structure of Francisella novicida Cas9 in complex with sgRNA and target DNA (TGA PAM)
Summary for 5B2P
| Entry DOI | 10.2210/pdb5b2p/pdb |
| Related | 5B2O 5B2Q |
| Descriptor | CRISPR-associated endonuclease Cas9, ACETATE ION, Guide RNA, ... (11 entities in total) |
| Functional Keywords | crispr-cas9, genome engineering, hydrolase-rna-dna complex, hydrolase/rna/dna |
| Biological source | Francisella tularensis subsp. novicida U112 More |
| Total number of polymer chains | 4 |
| Total formula weight | 235532.22 |
| Authors | Hirano, H.,Nishimasu, H.,Nakane, T.,Ishitani, R.,Nureki, O. (deposition date: 2016-02-01, release date: 2016-03-02, Last modification date: 2024-03-20) |
| Primary citation | Hirano, H.,Gootenberg, J.S.,Horii, T.,Abudayyeh, O.O.,Kimura, M.,Hsu, P.D.,Nakane, T.,Ishitani, R.,Hatada, I.,Zhang, F.,Nishimasu, H.,Nureki, O. Structure and Engineering of Francisella novicida Cas9 Cell, 164:950-961, 2016 Cited by PubMed Abstract: The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets complementary to the guide RNA and has been applied to programmable genome editing. Cas9-mediated cleavage requires a protospacer adjacent motif (PAM) juxtaposed with the DNA target sequence, thus constricting the range of targetable sites. Here, we report the 1.7 Å resolution crystal structures of Cas9 from Francisella novicida (FnCas9), one of the largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets. A structural comparison of FnCas9 with other Cas9 orthologs revealed striking conserved and divergent features among distantly related CRISPR-Cas9 systems. We found that FnCas9 recognizes the 5'-NGG-3' PAM, and used the structural information to create a variant that can recognize the more relaxed 5'-YG-3' PAM. Furthermore, we demonstrated that the FnCas9-ribonucleoprotein complex can be microinjected into mouse zygotes to edit endogenous sites with the 5'-YG-3' PAM, thus expanding the target space of the CRISPR-Cas9 toolbox. PubMed: 26875867DOI: 10.1016/j.cell.2016.01.039 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.7 Å) |
Structure validation
Download full validation report
