Table 1.

Cryo-EM data collection, refinement and validation statistics

	Al3Cas12f state I	Al3Cas12f state II	OsCas12f state I	OsCas12f state II	OsCas12f state III	RhCas12f
Data collection and processing
Magnification	150,000	150,000	150,000	150,000	150,000	105,000
Voltage (kV)	200	200	200	200	200	300
Electron exposure (e− per Å2)	50	50	50	50	50	80
Defocus range (μm)	−1.5 to −2.5	−1.5 to −2.5	−1.5 to −2.5	−1.5 to −2.5	−1.5 to −2.5	−1.5 to −2.5
Pixel size (Å)	0.94	0.94	0.94	0.94	0.94	0.83
Initial particle (no.)	2,039,247	2,039,247	223,671	223,671	223,671	875,624
Final particle (no.)	141,537	106,204	132,962	44,815	42,153	316,144
Map resolution (Å)	3.17	3.20	3.27	3.67	3.68	3.07
FSC threshold	0.143	0.143	0.143	0.143	0.143	0.143
Refinement
Initial model used	ModelAngelo	ModelAngelo	AlphaFold	9NZT	9NZT	AlphaFold
Model resolution (Å)	3.19	3.25	3.2	3.7	3.6	3.1
FSC threshold	0.143	0.143	0.143	0.143	0.143	0.143
Map sharpening B factor (Å2)	−109.1	−117.0	−127.7	−107.4	−104.3	−135.7
Model composition
Nonhydrogen atoms	9,025	11,066	7,928	9,260	9,448	7,397
Protein residues	694	889	598	750	728	389
Nucleotides	169	185	146	152	170	199
Ligands	N/A	N/A	Zn: 1	Zn: 1	Zn: 1	N/A
Mean B factors (Å2)
Protein	93.36	83.04	70.81	39.83	38.94	59.33
Nucleotides	164.30	117.10	94.92	64.73	67.14	151.79
Ligands	N/A	N/A	159.75	93.81	101.67	N/A
Root-mean-square deviations
Bond lengths (Å)	0.005	0.004	0.005	0.006	0.007	0.006
Bond angles (°)	0.883	0.843	1.035	1.216	1.250	1.116
Validation
MolProbity score	1.54	1.32	1.34	1.53	1.50	1.35
Clashscore	5.46	5.86	4.19	5.79	7.06	6.39
Poor rotamers (%)	0.61	0.53	0.77	1.09	0.48	0.28
Ramachandran plot
Favored (%)	97.38	98.40	97.29	96.88	97.47	98.70
Allowed (%)	2.62	1.60	2.71	3.12	2.53	1.30
Disallowed (%)	0	0	0	0	0	0
Protein Data Bank	9NZO	9NZR	9NZT	9NZQ	9NZS	9NZP
EM Data Bank	EMD-49954	EMD-49957	EMD-49959	EMD-49956	EMD-49958	EMD-49955