REL 2023-03-09 2023-04-11 2023-04-11 0000-0002-3866-2742 Fatma Esra Demircioglu Broad Institute Cambridge United States 02142 0000-0002-3866-2742 Fatma Esra Demircioglu Broad Institute Cambridge United States 02142 Strecker J Demircioglu FE Li D Faure G Wilkinson ME Macrae RK Zhang F National Institutes of Health/National Eye Institute (NIH/NEI) 1DP1-HL141201 United States National Institutes of Health/National Heart, Lung, and Blood Institute (NIH/NHLBI) 2R01HG009761-05 United States 5.7 10.6019/EMPIAR-11466 EMDB molecule EMD-28064 EMD-28070 EMD-28065 EMD-28071 EMD-28072 EMD-28073 Strecker J Demircioglu FE Li D Faure G Wilkinson ME Gootenberg JS Abudayyeh OO Nishimasu H Macrae RK Zhang F Science United States 6622 378 874 881 2022 English 10.1126/science.add7450 36423276 Unaligned movies of the Cas7-11-crRNA-Csx29 complex (Dataset_inactive_CASP) /data/EMPIAR_CASP/Dataset_inactive_CASP micrographs - multiframe TIFF TIFF 16553 31 UNSIGNED BYTE 5760 0.6632 4092 0.6632

Unaligned, non-gain-normalised, 31-frame movies from a Bioquantum-K3 camera recorded in super-resolution mode with twofold binning at 130,000× magnification. 16553 movies were collected with EPU on the MIT.nano Titan Krios G3i operated at 300 kV. Each micrograph accumulates 40 electrons per square Angstrom over 31 frames (0.69 s). Nominal pixel size is 0.6632 Å/pixel. Gain reference image for the dataset is attached. Final refinement star files produced from focused refinements for each reconstruction deposited on EMDB are also attached: EMD-28064 at 3.0 Å resolution, and EMD-28070 at 3.2 Å resolution. Data was processed in RELION 4.0. Further details about the dataset: Grid: Quantifoil R1.2/1.3 on 300 copper mesh C2 aperture: 50 um Spot: 4 Beam diameter: 0.75 um EF slit: 20 eV Acquisition pattern: 4 shots/hole x 12 um AFIS Physical Pixel: 0.6632 A/pix SuperRes pixel: 0.3316 A/pix Exposure rate: 25.53 e/pix/s Exposure time: 0.69 s Total exposure: 40.07 e/A2 Frames: 31 Defocus: 0.5 - 2 um

Unaligned movies of the Cas7-11-crRNA-target RNA-Csx29-Csx30 complex (Dataset_active_CASP) /data/EMPIAR_CASP/Dataset_active_CASP micrographs - multiframe TIFF TIFF 32890 31 UNSIGNED BYTE 5760 0.6632 4092 0.6632

Unaligned, non-gain-normalised, 31-frame movies from a Bioquantum-K3 camera recorded in super-resolution mode with twofold binning at 130,000× magnification. 32890 movies were collected with EPU on the MIT.nano Titan Krios G3i operated at 300 kV. Each micrograph accumulates 40 electrons per square Angstrom over 31 frames (0.69 s). Nominal pixel size is 0.6632 Å/pixel. Gain reference image for the dataset is attached. Final refinement star files produced from focused refinements for each reconstruction deposited on EMDB are also attached: EMD-28065 at 2.5 Å resolution, EMD-28071 at 2.8 Å resolution, EMD-28072 at 2.6 Å resolution, and EMD-28073 at 2.7 Å resolution. Only one-third of the dataset was used during processing in RELION 4.0. Further details about the dataset: Grid: Quantifoil R1.2/1.3 on 300 copper mesh C2 aperture: 50 um Spot: 4 Beam diameter: 0.75 um EF slit: 20 eV Acquisition pattern: 4 shots/hole x 12 um AFIS Physical Pixel: 0.6632 A/pix SuperRes pixel: 0.3316 A/pix Exposure rate: 25.53 e/pix/s Exposure time: 0.69 s Total exposure: 40.07 e/A2 Frames: 31 Defocus: 0.5 - 2 um