› 5nfv

X-ray diffraction
2.5Å resolution

Crystal structure of catalytically inactive FnCas12 mutant bound to an R-loop structure containing a pre-crRNA mimic and full-length DNA target

Released:
DOI: 10.2210/pdb5nfv/pdb
PDB entry 5nfv coloured by chain, front view.
PDB entry 5nfv coloured by chain, side view.
PDB entry 5nfv coloured by chain, top view.
Source organisms:
Primary publication:
Structural Basis for Guide RNA Processing and Seed-Dependent DNA Targeting by CRISPR-Cas12a.
Swarts DC, van der Oost J, Jinek M
Mol Cell 66 221-233.e4 (2017)
PMID: 28431230

Function and Biology Details

Reactions catalysed: 
Endonucleolytic cleavage to 5'-phosphodinucleotide and 5'-phosphooligonucleotide end-products
RNA = a 5'-hydroxy-ribonucleotide + n nucleoside-2',3'-cyclophosphates
Biochemical function:
Biological process:
Cellular component:
  • not assigned
Sequence domains:

Structure analysis Details

Assembly composition:
hetero tetramer (preferred)
Assembly name:
PDBe Complex ID:
PDB-CPX-106187 (preferred)
Entry contents:
1 distinct polypeptide molecule
2 distinct DNA molecules
1 distinct DNA/RNA hybrid molecule
Macromolecules (4 distinct):
CRISPR-associated endonuclease Cas12a Chain: A
Molecule details ›
Chain: A
Length: 1302 amino acids
Theoretical weight: 152.2 KDa
Source organism: Francisella tularensis subsp. novicida U112
Expression system: Escherichia coli BL21(DE3)
UniProt:
  • Canonical: A0Q7Q2 (Residues: 2-1300; Coverage: 100%)
Gene names: FTN_1397, cas12a, cpf1
Sequence domains:
DNA target strand Chain: C
Molecule details ›
Chain: C
Length: 38 nucleotides
Theoretical weight: 11.67 KDa
Source organism: synthetic construct
Expression system: Not provided
DNA non-target strand Chain: D
Molecule details ›
Chain: D
Length: 38 nucleotides
Theoretical weight: 11.62 KDa
Source organism: synthetic construct
Expression system: Not provided
pre-crRNA Chain: B
Molecule details ›
Chain: B
Length: 46 nucleotides
Theoretical weight: 14.66 KDa
Source organism: synthetic construct
Expression system: Not provided

Ligands and Environments

2 bound ligands:
Ligand MG 3 x MG
Ligand B3P 1 x B3P
No modified residues

Experiments and Validation Details

Entry percentile scores
X-ray source: SLS BEAMLINE X06DA
Spacegroup: P21
Unit cell:
a: 82.421Å b: 143.703Å c: 84.628Å
α: 90° β: 94.16° γ: 90°
R-values:
R R work R free
0.207 0.205 0.238
Expression systems:
  • Escherichia coli BL21(DE3)
  • Not provided

Quick links

Citations

49 review citations

Diversity, classification and evolution of CRISPR-Cas systems.
Koonin et al. (2017)
48 more

10 mentions without citation

The Revolution Continues: Newly Discovered Systems Expand the CRISPR-Cas Toolkit.
Murugan et al. (2017)
9 more