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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-22258 | |||||||||
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| Title | Cryo-EM structure of Cas12g ternary complex | |||||||||
Map data | unsharpened | |||||||||
Sample |
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Keywords | CRISPR / HYDROLASE-RNA complex | |||||||||
| Biological species | metagenome (others) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 4.8 Å | |||||||||
Authors | Chang L / Li Z / Zhang H | |||||||||
Citation | Journal: Nat Chem Biol / Year: 2021Title: Cryo-EM structure of the RNA-guided ribonuclease Cas12g. Authors: Zhuang Li / Heng Zhang / Renjian Xiao / Ruijie Han / Leifu Chang / ![]() Abstract: Cas12g, the type V-G CRISPR-Cas effector, is an RNA-guided ribonuclease that targets single-stranded RNA substrate. The CRISPR-Cas12g system offers a potential platform for transcriptome engineering ...Cas12g, the type V-G CRISPR-Cas effector, is an RNA-guided ribonuclease that targets single-stranded RNA substrate. The CRISPR-Cas12g system offers a potential platform for transcriptome engineering and diagnostic applications. We determined the structures of Cas12g-guide RNA complexes in the absence and presence of target RNA by cryo-EM to a resolution of 3.1 Å and 4.8 Å, respectively. Cas12g adopts a bilobed structure with miniature REC2 and Nuc domains, whereas the guide RNAs fold into a flipped 'F' shape, which is primarily recognized by the REC lobe. Target RNA and the CRISPR RNA (crRNA) guide form a duplex that inserts into the central cavity between the REC and NUC lobes, inducing conformational changes in both lobes to activate Cas12g. The structural insights would facilitate the development of Cas12g-based applications. | |||||||||
| History |
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Structure visualization
| Movie |
Movie viewer |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_22258.map.gz | 18.8 MB | EMDB map data format | |
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| Header (meta data) | emd-22258-v30.xml emd-22258.xml | 17 KB 17 KB | Display Display | EMDB header |
| Images | emd_22258.png | 57.2 KB | ||
| Filedesc metadata | emd-22258.cif.gz | 6.7 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-22258 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-22258 | HTTPS FTP |
-Validation report
| Summary document | emd_22258_validation.pdf.gz | 388.8 KB | Display | EMDB validaton report |
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| Full document | emd_22258_full_validation.pdf.gz | 388.5 KB | Display | |
| Data in XML | emd_22258_validation.xml.gz | 5.6 KB | Display | |
| Data in CIF | emd_22258_validation.cif.gz | 6.4 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22258 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22258 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6xmgMC ![]() 6xmfC C: citing same article ( M: atomic model generated by this map |
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| Similar structure data |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_22258.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | unsharpened | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.05 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : CRISP-Cas complex
| Entire | Name: CRISP-Cas complex |
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| Components |
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-Supramolecule #1: CRISP-Cas complex
| Supramolecule | Name: CRISP-Cas complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#3 |
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| Source (natural) | Organism: metagenome (others) |
-Macromolecule #1: CRISPR-Cas
| Macromolecule | Name: CRISPR-Cas / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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| Source (natural) | Organism: metagenome (others) |
| Molecular weight | Theoretical: 89.89168 KDa |
| Recombinant expression | Organism: ![]() |
| Sequence | String: MAQASSTPAV SPRPRPRYRE ERTLVRKLLP RPGQSKQEFR ENVKKLRKAF LQFNADVSGV CQWAIQFRPR YGKPAEPTET FWKFFLEPE TSLPPNDSRS PEFRRLQAFE AAAGINGAAA LDDPAFTNEL RDSILAVASR PKTKEAQRLF SRLKDYQPAH R MILAKVAA ...String: MAQASSTPAV SPRPRPRYRE ERTLVRKLLP RPGQSKQEFR ENVKKLRKAF LQFNADVSGV CQWAIQFRPR YGKPAEPTET FWKFFLEPE TSLPPNDSRS PEFRRLQAFE AAAGINGAAA LDDPAFTNEL RDSILAVASR PKTKEAQRLF SRLKDYQPAH R MILAKVAA EWIESRYRRA HQNWERNYEE WKKEKQEWEQ NHPELTPEIR EAFNQIFQQL EVKEKRVRIC PAARLLQNKD NC QYAGKNK HSVLCNQFNE FKKNHLQGKA IKFFYKDAEK YLRCGLQSLK PNVQGPFRED WNKYLRYMNL KEETLRGKNG GRL PHCKNL GQECEFNPHT ALCKQYQQQL SSRPDLVQHD ELYRKWRREY WREPRKPVFR YPSVKRHSIA KIFGENYFQA DFKN SVVGL RLDSMPAGQY LEFAFAPWPR NYRPQPGETE ISSVHLHFVG TRPRIGFRFR VPHKRSRFDC TQEELDELRS RTFPR KAQD QKFLEAARKR LLETFPGNAE QELRLLAVDL GTDSARAAFF IGKTFQQAFP LKIVKIEKLY EQWPNQKQAG DRRDAS SKQ PRPGLSRDHV GRHLQKMRAQ ASEIAQKRQE LTGTPAPETT TDQAAKKATL QPFDLRGLTV HTARMIRDWA RLNARQI IQ LAEENQVDLI VLESLRGFRP PGYENLDQEK KRRVAFFAHG RIRRKVTEKA VERGMRVVTV PYLASSKVCA ECRKKQKD N KQWEKNKKRG LFKCEGCGSQ AQVDENAARV LGRVFWGEIE LPTAIP |
-Macromolecule #2: RNA (130-MER)
| Macromolecule | Name: RNA (130-MER) / type: rna / ID: 2 / Number of copies: 1 |
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| Source (natural) | Organism: metagenome (others) |
| Molecular weight | Theoretical: 43.777043 KDa |
| Sequence | String: GGGAUGCUUA CUUAGUCAUC UGGUUGGCAA ACCUCCGCGG ACCUUCGGGA CCAAUGGAGA GGAACCCAGC CGAGAAGCAU CGAGCCGGU AAAUGUUUAC CGGCUCUGAC ACCAACUGUG AUAAACUACC GCAUUAA |
-Macromolecule #3: RNA (5'-R(P*UP*UP*AP*AP*UP*GP*CP*GP*GP*UP*AP*GP*UP*UP*UP*AP*UP*CP...
| Macromolecule | Name: RNA (5'-R(P*UP*UP*AP*AP*UP*GP*CP*GP*GP*UP*AP*GP*UP*UP*UP*AP*UP*CP*AP*CP*AP*GP*UP*U)-3') type: rna / ID: 3 / Number of copies: 1 |
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| Source (natural) | Organism: metagenome (others) |
| Molecular weight | Theoretical: 7.633511 KDa |
| Sequence | String: UUAAUGCGGU AGUUUAUCAC AGUU |
-Macromolecule #4: ZINC ION
| Macromolecule | Name: ZINC ION / type: ligand / ID: 4 / Number of copies: 1 / Formula: ZN |
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| Molecular weight | Theoretical: 65.409 Da |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.5 |
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| Grid | Details: unspecified |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 1.35 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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