Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter to the Editor
  • Published:

Structure and genome editing activity of the novel CRISPR-Cas12o1 effector

Cell Research volume 35, pages 145–148 (2025)Cite this article

Subjects

Access through your institution
Buy or subscribe

Dear Editor,

Class 2 type V CRISPR-Cas12 effectors exhibit tremendous diversity, utilizing either single crRNAs or dual RNA guides to target double-stranded DNA (dsDNA), single-stranded DNA (ssDNA) or RNA. For dsDNA targets, Cas12 recognizes different PAM sequences and generates sticky ends on the target dsDNA.1 This characteristic enhances the efficiency of homology-directed repair2 and causes less off-target effects compared to SpCas9.3 Recent studies have identified a series of Cas12 nucleases, ranging from Cas12a to Cas12n,1,4 thereby greatly expanding the gene editing toolkit. However, among the reported Cas12 proteins, the compact ones typically show limited gene editing efficiency and associate with long non-coding RNA guides or dual RNA (crRNA and tracrRNA) guides, which may compensate for the absence of certain Cas12 domains and enhance endonuclease activity and editing efficiency.5

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Structures and engineering of the novel Cas12o1 nucleases.

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

  1. Yan, W. X. et al. Science 363, 88–91 (2019).

    Article  PubMed  CAS  Google Scholar 

  2. Moreno-Mateos, M. A. et al. Nat. Commun. 8, 2024 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  3. Kleinstiver, B. P. et al. Nat. Biotechnol. 34, 869–874 (2016).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Chen, W. et al. Mol. Cell 83, 2768–2780.e6 (2023).

    Article  PubMed  CAS  Google Scholar 

  5. Sun, A. et al. Cell Res. 33, 229–244 (2023).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Markowitz, V. M. et al. Nucleic Acids Res. 40, D123–D129 (2012).

    Article  PubMed  CAS  Google Scholar 

  7. Yang, H., Gao, P., Rajashankar, K. R. & Patel, D. J. Cell 167, 1814–1828.e12 (2016).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  8. Kong, X. et al. Nat. Commun. 14, 2046 (2023).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  9. Walton, R. T., Christie, K. A., Whittaker, M. N. & Kleinstiver, B. P. Science 368, 290–296 (2020).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

Download references

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.

Author information

Author notes

  1. These authors contributed equally: Zhiqiang Duan, Xi Zhang, Jun-Tao Zhang, Xingkun Ji.

Authors and Affiliations

  1. Institute of Advanced Biotechnology and School of Medicine, Southern University of Science and Technology, Shenzhen, Guangdong, China

    Zhiqiang Duan, Xi Zhang, Xingkun Ji & Jian-Kang Zhu

  2. Department of Biochemistry, School of Medicine, Southern University of Science and Technology, Shenzhen, Guangdong, China

    Jun-Tao Zhang & Ning Jia

  3. Bellagen Biotechnology Co. Ltd., Jinan, Shandong, China

    Ruiheng Liu, Ying Chen, Shanshan Li, Nannan Jia, Huizhi Gao & Yu Xin

  4. Shenzhen Key Laboratory of Cell Microenvironment, Guangdong Provincial Key Laboratory of Cell Microenvironment and Disease Research, Southern University of Science and Technology, Shenzhen, Guangdong, China

    Ning Jia

  5. Key University Laboratory of Metabolism and Health of Guangdong, Institute for Biological Electron Microscopy, Southern University of Science and Technology, Shenzhen, Guangdong, China

    Ning Jia

  6. Center for Advanced Bioindustry Technologies, Chinese Academy of Agricultural Sciences, Beijing, China

    Jian-Kang Zhu

Authors
  1. Zhiqiang Duan
    View author publications

    Search author on:PubMed Google Scholar

  2. Xi Zhang
    View author publications

    Search author on:PubMed Google Scholar

  3. Jun-Tao Zhang
    View author publications

    Search author on:PubMed Google Scholar

  4. Xingkun Ji
    View author publications

    Search author on:PubMed Google Scholar

  5. Ruiheng Liu
    View author publications

    Search author on:PubMed Google Scholar

  6. Ying Chen
    View author publications

    Search author on:PubMed Google Scholar

  7. Shanshan Li
    View author publications

    Search author on:PubMed Google Scholar

  8. Nannan Jia
    View author publications

    Search author on:PubMed Google Scholar

  9. Huizhi Gao
    View author publications

    Search author on:PubMed Google Scholar

  10. Yu Xin
    View author publications

    Search author on:PubMed Google Scholar

  11. Ning Jia
    View author publications

    Search author on:PubMed Google Scholar

  12. Jian-Kang Zhu
    View author publications

    Search author on:PubMed Google Scholar

Contributions

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.

Corresponding authors

Correspondence to Ning Jia or Jian-Kang Zhu.

Ethics declarations

Competing interests

Z.D. and Y.C. are inventors on a patent application related to this work submitted by Shandong BellaGen Biotechnology Co., Ltd.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/s41422-024-01050-y

This article is cited by

Search

Advanced search

Quick links