|Entry||Database: EMDB / ID: 6777|
|Title||Electron cryo-microscopy structure of LbuCas13a-crRNA binary complex|
|Sample||LbuCas13a-crRNA binary complex|
|Source||Leptotrichia buccalis / bacteria / レプトトリキア・ブカリス|
|Map data||Electron cryo-microscopy structure of LbuCas13a-crRNA binary complex|
|Method||single particle reconstruction, at 3.2 Å resolution|
|Authors||Zhang X / Wang Y|
|Citation||Cell, 2017, 170, 714-726.e10|
|Validation Report||PDB-ID: 5xwy|
SummaryFull reportAbout validation report
|Date||Deposition: Jun 30, 2017 / Header (metadata) release: Sep 13, 2017 / Map release: Sep 13, 2017 / Last update: Sep 13, 2017|
Downloads & links
|File||emd_6777.map.gz (map file in CCP4 format, 32001 KB)|
|Projections & slices|
Images are generated by Spider package.
|Voxel size||X=Y=Z: 0.82 Å|
CCP4 map header:
-Entire LbuCas13a-crRNA binary complex
|Entire||Name: LbuCas13a-crRNA binary complex / Number of components: 3|
-Component #1: protein, LbuCas13a-crRNA binary complex
|Protein||Name: LbuCas13a-crRNA binary complex / Recombinant expression: No|
|Source||Species: Leptotrichia buccalis / bacteria / レプトトリキア・ブカリス|
Strain: ATCC 14201
|Source (engineered)||Expression System: Escherichia coli / bacteria / エシェリキア・コリ, 大腸菌 /|
-Component #2: protein, A type VI-A CRISPR-Cas RNA-guided RNA ribonuclease, Cas13a
|Protein||Name: A type VI-A CRISPR-Cas RNA-guided RNA ribonuclease, Cas13a|
Recombinant expression: No
|Mass||Theoretical: 138.555156 kDa|
|Source (engineered)||Expression System: Leptotrichia buccalis (strain atcc 14201 / dsm 1135 / jcm 12969 / nctc 10249 / c-1013-b) / bacteria|
Strain: ATCC 14201 / DSM 1135 / JCM 12969 / NCTC 10249 / C-1013-b
-Component #3: nucleic-acid, RNA (59-MER)
|Nucleic-acid||Name: RNA (59-MER) / Class: RNA / Structure: OTHER / Synthetic: No|
GGACCACCCC AAAAAUGAAG GGGACUAAAA CACAAAUCUA UCUGAAUAAA CUCUUCUUC
|Mass||Theoretical: 18.869348 kDa|
|Sample solution||pH: 7.5|
|Vitrification||Cryogen name: ETHANE|
-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: 60 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 (4k x 4k)|
|Processing||Method: single particle reconstruction / Number of projections: 238758|
|3D reconstruction||Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF|
-Atomic model buiding
-Oct 4, 2017. Three pioneers of this field were awarded Nobel Prize in Chemistry 2017
Three pioneers of this field were awarded Nobel Prize in Chemistry 2017
- Jacques Dubochet (University of Lausanne, Switzerland) is a pioneer of ice-embedding method of EM specimen (as known as cryo-EM), Most of 3DEM structures in EMDB and PDB are obtained using his method.
- Joachim Frank (Columbia University, New York, USA) is a pioneer of single particle reconstruction, which is the most used reconstruction method for 3DEM structures in EMDB and EM entries in PDB. And also, he is a develper of Spider, which is one of the most famous software in this field, and is used for some EM Navigor data (e.g. map projection/slice images).
- Richard Henderson (MRC Laboratory of Molecular Biology, Cambridge, UK) was determined the first biomolecule structure by EM. The first EM entry in PDB, PDB-1brd is determinedby him.
External links: The 2017 Nobel Prize in Chemistry - Press Release
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