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