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Biology and applications of CRISPR–Cas12 and transposon-associated homologs

Nature Biotechnology volume 42, pages 1807–1821 (2024)Cite this article

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Abstract

CRISPR-associated Cas12 proteins are a highly variable collection of nucleic acid-targeting proteins. All Cas12 variants use RNA guides and a single nuclease domain to target complementary DNA or, in rare cases, RNA. The high variability of Cas12 effectors can be explained by a series of independent evolution events from different transposon-associated TnpB-like ancestors. Despite basic structural and functional similarities, this has resulted in unprecedented variation of the Cas12 effector proteins in terms of size, domain composition, guide structure, target identity and interference strategy. In this Review, we compare the unique molecular features of natural and engineered Cas12 and TnpB variants. Furthermore, we provide an overview of established genome editing and diagnostic applications and discuss potential future directions.

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Fig. 1: Evolution of OMEGA nucleases to Cas9 and Cas12.
Fig. 2: Size range of OMEGA and type V effector proteins and associated RNAs.
Fig. 3: Different activities of Cas12 variants.
Fig. 4: Cas12a-based BEs and PEs.

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Acknowledgements

This research is supported by research grants of the Dutch Research Council (NWO; BBOL-737.016.005, Spinoza SPI 93–537 and Gravitation 024.003.019) and the European Research Council (ERC-AdG-834279).

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Author notes

  1. These authors contributed equally: Wen Y. Wu, Belén Adiego-Pérez.

Authors and Affiliations

  1. Laboratory of Microbiology, Wageningen University & Research, Wageningen, the Netherlands

    Wen Y. Wu, Belén Adiego-Pérez & John van der Oost

  2. Department of Chemical and Pharmaceutical Biology, Groningen Research Institute of Pharmacy, University of Groningen, Groningen, the Netherlands

    Wen Y. Wu

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W.Y.W., B.A.-P. and J.v.d.O. conceptualized the review and contributed to the writing of the manuscript. W.Y.W. and B.A.-P. prepared the visual elements, including figure and table design.

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Correspondence to Wen Y. Wu or John van der Oost.

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Competing interests

Several CRISPR–Cas-related patent applications have been filed related to this work, with J.v.d.O. and W.Y.W. as inventors. J.v.d.O. is cofounder and scientific adviser of NTrans Technologies and adviser of Scope Biosciences and Hudson River Biotechnology. B.A-P. declares no competing interests.

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Nature Biotechnology thanks Chase Beisel and Haoyi Wang for their contribution to the peer review of this work.

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Wu, W.Y., Adiego-Pérez, B. & van der Oost, J. Biology and applications of CRISPR–Cas12 and transposon-associated homologs. Nat Biotechnol 42, 1807–1821 (2024). https://doi.org/10.1038/s41587-024-02485-9

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