8ywh

Cryo-EM structure of Compact SaCas9-RNA-DNA ternary complex

spinqualitylabelsall models 8ywh, resolution 2.97Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image   Print3D Structural highlights 8ywh is a 4 chain structure with sequence from Staphylococcus aureus and Synthetic construct. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance. Method: Electron Microscopy, Resolution 2.97Å Resources: FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT Publication Abstract from PubMed Proteins with multiple domains play pivotal roles in various biological processes, necessitating a thorough understanding of their structural stability and functional interplay. Here, a structure-guided protein engineering approach is proposed to develop thermostable Cas9 (CRISPR-associated protein 9) variant for CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) interference applications. By employing thermodynamic analysis, combining distance mapping and molecular dynamics simulations, deletable domains are identified to enhance stability while preserving the DNA recognition function of Cas9. The resulting engineered Cas9, termed small and dead form Cas9, exhibits improved thermostability and maintains target DNA recognition function. Cryo-electron microscopy analysis reveals structural integrity with reduced atomic density in the deleted domain. Fusion with functional elements enables intracellular delivery and nuclear localization, demonstrating efficient gene suppression in diverse cell types. Direct delivery in the mouse brain shows enhanced knockdown efficiency, highlighting the potential of structure-guided engineering to develop functional CRISPR systems tailored for specific applications. This study underscores the significance of integrating computational and experimental approaches for protein engineering, offering insights into designing tailored molecular tools for precise biological interventions. Structure-Guided Engineering of Thermodynamically Enhanced SaCas9 for Improved Gene Suppression.,Kang ES, Kim NH, Lim HK, Jeon H, Han K, No YH, Kim K, Khaleel ZH, Shin D, Eom K, Nam J, Lee BS, Kim HJ, Suh M, Lee J, Thach TT, Hyun J, Kim YH Adv Mater. 2024 Jun 22:e2404680. doi: 10.1002/adma.202404680. PMID:38944889 From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.

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