4OO8
Crystal structure of Streptococcus pyogenes Cas9 in complex with guide RNA and target DNA
Experimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | SPRING-8 BEAMLINE BL32XU |
Synchrotron site | SPring-8 |
Beamline | BL32XU |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2013-11-12 |
Detector | RAYONIX MX-225 |
Wavelength(s) | 1.000 |
Spacegroup name | P 1 |
Unit cell lengths | 76.713, 105.694, 126.817 |
Unit cell angles | 97.68, 98.43, 100.31 |
Refinement procedure
Resolution | 46.701 - 2.500 |
R-factor | 0.2235 |
Rwork | 0.222 |
R-free | 0.25360 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | SeMet-labelled Cas9 |
RMSD bond length | 0.002 |
RMSD bond angle | 0.526 |
Data reduction software | XDS |
Data scaling software | XSCALE |
Phasing software | MOLREP |
Refinement software | PHENIX (1.8.3_1479) |
Data quality characteristics
Overall | Inner shell | Outer shell | |
Low resolution limit [Å] | 46.883 | 2.650 | |
High resolution limit [Å] | 2.500 | 7.410 | 2.500 |
Rmerge | 0.063 | 1.100 | 1.100 |
Rmeas | 0.019 | 1.134 | |
Number of reflections | 130485 | 4896 | 20643 |
<I/σ(I)> | 25.27 | 87.06 | 2.1 |
Completeness [%] | 98.2 | 96.4 | 96.2 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, HANGING DROP | 8 | 293 | 12% PEG 3350, 100mM Tris-HCl, 200mM ammonium acetate, 150mM NaCl, 100mM NDSB-256, pH 8.0, vapor diffusion, hanging drop, temperature 293K |