REL 2025-08-11 2025-11-05 2025-11-05 Cas1-2/3 integration complex, 32 bp foreign DNA without PAM, CRISPR leader-repeat-spacer DNA fragment 0000-0001-9836-3827 William Henriques Department of Microbiology and Cell Biology, Montana State University 128 Cooley Laboratory Bozeman Montana United States 59717 0000-0001-9297-5304 Blake Wiedenheft Department of Microbiology and Cellular Biology, Montana State University, Bozeman, MT 128 Cooley Laboratory Bozeman Montana United States 59717 Henriques WS Bowman J Hall LN Gauvin CC Wei H Kuang H Zimanyi CM Eng ET Santiago-Frangos A Wiedenheft B 6.1 10.6019/EMPIAR-12922 EMDB molecule EMD-71114 EMD-71115 EMD-71116 Henriques WS Bowman J Hall LN Gauvin CC Wei H Kuang H Zimanyi CM Eng ET Santiago-Frangos A Wiedenheft B Structures reveal how the Cas1-2/3 integrase captures, delivers, and integrates foreign DNA into CRISPR loci bioRxiv United States 2025 English 10.1101/2025.06.10.658980 40661357
The datasets for both the trimmed and PAM-containing integration complexes were collected on Montana State University's Talos Arctica transmission electron microscope (Thermo Fisher Scientific), with a field emission gun operating at an accelerating voltage of 200 keV using parallel illumination conditions. Movies were acquired using a Gatan K3 direct electron detector, operated in electron counting mode targeting a total electron exposure of 65.67 e-/Å2 over 50 frames (5.992 second exposure, 0.12 second frame time). The SerialEM data collection software was used to collect micrographs at 36,000x nominal magnification (1.152 Å/pixel at the specimen level) over a defocus range of -0.5 µm to -2.0 µm. Stage movement was used to target the center of four 2.0 µm holes for focusing, and image shift was used to acquire high magnification images in the center of each of the holes.
 Henriques WS Bowman J Hall LN Gauvin CC Wei H Kuang H Zimanyi CM Eng ET Santiago-Frangos A Wiedenheft B Structures reveal how the Cas1-2/3 integrase captures, delivers, and integrates foreign DNA into CRISPR loci Structure (London, England : 1993) Structure 2025 English 10.1016/j.str.2025.09.007 41072406 Unaligned multi-frame micrographs of the Cas1-2/3 heterohexameric complex bound to a fragment of dsDNA without a PAM and to a dsDNA fragment containing the CRISPR leader, repeat, and spacer /data/rawdata1-gr1 micrographs - multiframe TIFF TIFF 4000 50 1 50 UNSIGNED BYTE 5760 1.152 4092 1.152
The micrographs are unaligned, gain-corrected movies.
data/rawdata1-gr1/gr1*.tif Unaligned multi-frame micrographs of the Cas1-2/3 heterohexameric complex bound to a fragment of dsDNA without a PAM and to a dsDNA fragment containing the CRISPR leader, repeat, and spacer /data/rawdata2-gr1 micrographs - multiframe TIFF TIFF 3666 50 1 50 UNSIGNED BYTE 5760 1.152 4092 1.152
The micrographs are unaligned, gain-corrected movies.
data/rawdata2-gr1/gr1*.tif