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Open data
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Basic information
| Entry | Database: PDB / ID: 9lcz | |||||||||||||||||||||
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| Title | Inactivate TOD6 with GC DNA substrate | |||||||||||||||||||||
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Keywords | DE NOVO PROTEIN / De novo design / DNA-binding TALE domain / deaminase(Ddd_Ss) / orienting domain | |||||||||||||||||||||
| Function / homology | DNA / DNA (> 10) Function and homology information | |||||||||||||||||||||
| Biological species | Xanthomonas (bacteria)synthetic construct (others) | |||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.93 Å | |||||||||||||||||||||
Authors | Lv, X.C. / Mi, L. / Lu, P.L. | |||||||||||||||||||||
| Funding support | China, 2items
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Citation | Journal: Nat Struct Mol Biol / Year: 2025Title: Computational design of a high-precision mitochondrial DNA cytosine base editor. Authors: Li Mi / Yu-Xuan Li / Xinchen Lv / Zi-Li Wan / Xu Liu / Kairan Zhang / Huican Li / Yue Yao / Leping Zhang / Zhe Xu / Xingyu Zhuang / Kunqian Ji / Min Jiang / Yangming Wang / Peilong Lu / ![]() Abstract: Bystander editing remains a major limitation of current base editors, hindering their precision and therapeutic potential. Here, we present a de novo protein design strategy that creates a ...Bystander editing remains a major limitation of current base editors, hindering their precision and therapeutic potential. Here, we present a de novo protein design strategy that creates a structurally rigid interface between a DNA-binding TALE domain and a cytosine deaminase, forming a unified editing module termed TALE-oriented deaminase (TOD). Cryo-EM analysis of TOD-DNA complexes confirms that this precise spatial architecture tightly restricts the deaminase activity window, thereby minimizing unwanted deamination. To further enhance editing specificity, we develop a split version, termed DdCBE-TOD, which virtually eliminates off-target editing. As a proof of concept, we apply DdCBE-TOD to generate a mitochondrial disease mouse model and to correct a pathogenic mutation associated with MERRF syndrome in patient-derived cells, achieving single-nucleotide precision. This work introduces a generalizable and computationally guided approach for ultra-precise base editing, offering a promising platform for both mechanistic studies and therapeutic correction of single-nucleotide mutations. | |||||||||||||||||||||
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 9lcz.cif.gz | 194.1 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb9lcz.ent.gz | 147.5 KB | Display | PDB format |
| PDBx/mmJSON format | 9lcz.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 9lcz_validation.pdf.gz | 360.9 KB | Display | wwPDB validaton report |
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| Full document | 9lcz_full_validation.pdf.gz | 367.7 KB | Display | |
| Data in XML | 9lcz_validation.xml.gz | 16.2 KB | Display | |
| Data in CIF | 9lcz_validation.cif.gz | 26.3 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/lc/9lcz ftp://data.pdbj.org/pub/pdb/validation_reports/lc/9lcz | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 62997MC ![]() 9lcxC ![]() 9lcyC ![]() 9ld0C ![]() 9ld1C C: citing same article ( M: map data used to model this data |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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| 1 |
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Components
| #1: Protein | Mass: 93232.461 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Xanthomonas (bacteria) / Production host: ![]() |
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| #2: DNA chain | Mass: 9473.098 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) |
| #3: DNA chain | Mass: 9593.171 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) synthetic construct (others) |
| #4: Chemical | ChemComp-ZN / |
| Has ligand of interest | Y |
| Has protein modification | N |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
| Component | Name: Inactivate TOD6 with GC DNA substrate / Type: COMPLEX / Entity ID: #1-#3 / Source: RECOMBINANT |
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| Source (natural) | Organism: Xanthomonas (bacteria) |
| Source (recombinant) | Organism: ![]() |
| Buffer solution | pH: 6 |
| Specimen | Conc.: 6 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: TFS KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2000 nm / Nominal defocus min: 1400 nm |
| Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||
| 3D reconstruction | Resolution: 2.93 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 964225 / Symmetry type: POINT | ||||||||||||||||
| Atomic model building | Protocol: AB INITIO MODEL |
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About Yorodumi




Xanthomonas (bacteria)
China, 2items
Citation








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FIELD EMISSION GUN