|Entry||Database: EMDB / ID: 8749|
|Title||cryo-EM structure of the Cas9-sgRNA-AcrIIA4 anti-CRISPR complex|
|Sample||the complex of Streptococcus progenies Cas9 with sgRNA and AcrIIA4|
|Source||Streptococcus pyogenes m1 gas / bacteria|
unidentified phage / virus
|Map data||Cas9- sgRNA-AcrIIA4 anti-CRISPR complex|
|Method||single particle reconstruction, at 3.9 Å resolution|
|Authors||Jiang F / Liu JJ|
|Citation||Sci Adv, 2017, 3, e1701620-e1701620|
|Validation Report||PDB-ID: 5vzl|
SummaryFull reportAbout validation report
|Date||Deposition: May 29, 2017 / Header (metadata) release: Jul 26, 2017 / Map release: Jul 26, 2017 / Last update: Jan 31, 2018|
Downloads & links
|File||emd_8749.map.gz (map file in CCP4 format, 67109 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 1.07 Å|
CCP4 map header:
+Entire the complex of Streptococcus progenies Cas9 with sgRNA and AcrIIA4
|Entire||Name: the complex of Streptococcus progenies Cas9 with sgRNA and AcrIIA4|
Number of components: 6
+Component #1: protein, the complex of Streptococcus progenies Cas9 with sgRNA a...
|Protein||Name: the complex of Streptococcus progenies Cas9 with sgRNA and AcrIIA4|
Recombinant expression: No
|Source||Species: Streptococcus pyogenes m1 gas / bacteria|
|Source (engineered)||Expression System: Escherichia coli / bacteria / /|
+Component #2: protein, Streptococcus pyogenes M1 GAS
|Protein||Name: Streptococcus pyogenes M1 GAS / Recombinant expression: No|
|Source||Species: unidentified phage / virus|
+Component #3: protein, unidentified phage
|Protein||Name: unidentified phage / Recombinant expression: No|
+Component #4: nucleic-acid, single guide RNA (116-MER)
|Nucleic-acid||Name: single guide RNA (116-MER) / Class: RNA / Structure: OTHER / Synthetic: No|
(GTP)GCGCAUAAA GAUGAGACGC GUUUUAGAGC UAUGCUGUUU UGAAAAAAAC AGCAUAGCAA GUUAAAAUAA GGCUAGUCCG UUAUCAACUU GAAAAAGUGG CACCGAGUCG GUGCUUCG
|Mass||Theoretical: 38.292645 kDa|
+Component #5: protein, CRISPR-associated endonuclease Cas9
|Protein||Name: CRISPR-associated endonuclease Cas9 / Recombinant expression: No|
|Mass||Theoretical: 158.772891 kDa|
|Source (engineered)||Expression System: Streptococcus pyogenes m1 gas / bacteria|
+Component #6: protein, phage anti-CRISPR AcrIIA4
|Protein||Name: phage anti-CRISPR AcrIIA4 / Recombinant expression: No|
|Mass||Theoretical: 10.182073 kDa|
|Source (engineered)||Expression System: unidentified phage / virus|
|Sample solution||Specimen conc.: 1.25 mg/ml / pH: 8|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Temperature: 281 K / Humidity: 100 %|
Details: blot for 4.5 seconds with force of 12 before plunging
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 46 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Cs: 0.01 mm / Imaging mode: BRIGHT FIELD / Defocus: 1000 - 3000 nm|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 (4k x 4k)|
|Processing||Method: single particle reconstruction / Number of projections: 185000|
|3D reconstruction||Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF|
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Three pioneers of this field were awarded Nobel Prize in Chemistry 2017
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