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Data availability
The cryo-EM density maps of the Cas12o1crRNA–dsDNA ternary complex and Cas12o1 (E100K)crRNA–dsDNA ternary complex were deposited in the Electron Microscopy Data Bank (EMDB) under the accession numbers EMD-38238 and EMD-38239. The corresponding atomic coordinates were deposited in the Protein Data Bank (PDB) with accession numbers 8XCA and 8XCC. Source data are provided in Supplementary information, Table S2. Primers and oligonucleotides used in this paper are provided in Supplementary information, Table S3.
References
Yan, W. X. et al. Science 363, 88–91 (2019).
Moreno-Mateos, M. A. et al. Nat. Commun. 8, 2024 (2017).
Kleinstiver, B. P. et al. Nat. Biotechnol. 34, 869–874 (2016).
Chen, W. et al. Mol. Cell 83, 2768–2780.e6 (2023).
Sun, A. et al. Cell Res. 33, 229–244 (2023).
Markowitz, V. M. et al. Nucleic Acids Res. 40, D123–D129 (2012).
Yang, H., Gao, P., Rajashankar, K. R. & Patel, D. J. Cell 167, 1814–1828.e12 (2016).
Kong, X. et al. Nat. Commun. 14, 2046 (2023).
Walton, R. T., Christie, K. A., Whittaker, M. N. & Kleinstiver, B. P. Science 368, 290–296 (2020).
Acknowledgements
This work was supported by Shandong BellaGen Biotechnology Co., Ltd., and by grants from the National Natural Science Foundation of China (32188102 to J.-K.Z. and 32270050 to Ning J.), the Guangdong and Shenzhen Natural Science Foundation (2024A1515010541 and JCYJ20220530114409022 to Ning J.), the Guangdong Provincial Science and Technology Innovation Council Grant (2017B030301018). Ning J. is an investigator of Southern University of Science and Technology (SUSTech) Institute for Biological Electron Microscopy. We thank the staff at SUSTech Cryo-EM Center for assistance with data collection on the SUSTech Titan KRIOS cryo-electron microscope.
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J.-K.Z. supervised the project. J.-K.Z., Ning J., and Z.D. designed the experiments and analyzed the data. Z.D. performed structure-based protein engineering, and provided materials and experimental advice for structural analysis. X.Z. conducted biochemical and structural studies, performed sample preparation and purification, and cryo-EM data collection. X.J. constructed the binary vectors for plant transformation, obtained regenerated plants and performed mammalian cell transfection. J.-T.Z. performed data processing, structure refinement and data analysis. S.L. built the phylogenetic tree and performed computational analyses for PAM depletion assays. R.L., Y.C., H.G., Y.X., and Nannan J. made constructs, performed gene editing experiments in mammalian cells and the in vitro cleavage assay of Cas12o1. Ning J., J.-K.Z. and J.-T.Z. wrote the manuscript with input from all authors. All authors read the manuscript and agreed to its publication.
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Z.D. and Y.C. are inventors on a patent application related to this work submitted by Shandong BellaGen Biotechnology Co., Ltd.
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Duan, Z., Zhang, X., Zhang, JT. et al. Structure and genome editing activity of the novel CRISPR-Cas12o1 effector. Cell Res 35, 145–148 (2025). https://doi.org/10.1038/s41422-024-01050-y
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DOI: https://doi.org/10.1038/s41422-024-01050-y
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