9KJ6
Crystal Structure of SpCas9 ternary complex, amino acids (1242-1263) replaced with Gly-Ser linker
Summary for 9KJ6
| Entry DOI | 10.2210/pdb9kj6/pdb |
| Descriptor | CRISPR-associated endonuclease Cas9/Csn1, single-guide RNA, Target Strand of double-stranded DNA, ... (4 entities in total) |
| Functional Keywords | crispr-cas9, dna binding protein |
| Biological source | Streptococcus pyogenes More |
| Total number of polymer chains | 4 |
| Total formula weight | 211170.04 |
| Authors | Kim, S.,Bae, J.,Choi, H.J. (deposition date: 2024-11-12, release date: 2025-11-12, Last modification date: 2026-05-27) |
| Primary citation | Kim, S.,Won, H.,Bae, J.,Kim, J.,Choi, J.,Richar, H.,Kim, Y.G.,Choi, H.J. Structural and functional insights into internal domain replacement in SpCas9 for protein engineering. Sci Rep, 15:41528-41528, 2025 Cited by PubMed Abstract: The CRISPR-Cas9 system has emerged as a powerful tool for precise genome editing, with ongoing research focused on enhancing its reliability and expanding its versatility. One effective strategy involves the integration of foreign functional domains into Cas9 to confer new capabilities. However, successful integration requires identification of insertion sites that preserve the protein's structural integrity and function. In this study, we identified a C-terminal region of Streptococcus pyogenes Cas9 (SpCas9), spanning residues 1242-1263, as a viable site for domain replacement. Structural and biochemical analyses of a SpCas9 variant lacking this region confirmed its dispensability for SpCas9 activity. As a proof of concept, we substituted this segment with the evolved E. coli tRNA adenosine deaminase (TadA), a key component of adenine base editors. Functional evaluation of this engineered SpCas9-TadA variant demonstrated deamination efficiency comparable to that of the ABE8e, with the potential to modulate the editing window through linker design. These results highlight the potential of targeted engineering of this region to develop more precise and versatile genome editing tools. PubMed: 41285890DOI: 10.1038/s41598-025-25367-9 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (3.59 Å) |
Structure validation
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