A miniature CRISPR-Cas10 enzyme confers immunity by inhibitory signalling

  • Nature. 2025 Oct 1. doi: 10.1038/s41586-025-09569-9.
Erin E Doherty  #  1  2  3 ,  Benjamin A Adler  #  1  2  3 ,  Peter H Yoon  1  2 ,  Kendall Hsieh  2 ,  Kenneth Loi  2 ,  Emily G Armbruster  4 ,  Arushi Lahiri  2 ,  Cydni S Bolling  1 ,  Xander E Wilcox  5 ,  Amogha Akkati  6 ,  Anthony T Iavarone  3 ,  Joe Pogliano  4 ,  Jennifer A Doudna  7  8  9  10  11  12  13
Affiliations
  • 1. Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA.
  • 2. Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA.
  • 3. California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, CA, USA.
  • 4. School of Biological Sciences, University of California San Diego, La Jolla, CA, USA.
  • 5. Department of Microbiology and Immunology, Cornell University, Ithaca, NY, USA.
  • 6. Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, CA, USA.
  • 7. Innovative Genomics Institute, University of California, Berkeley, Berkeley, CA, USA. [email protected].
  • 8. Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA, USA. [email protected].
  • 9. California Institute for Quantitative Biosciences (QB3), University of California, Berkeley, Berkeley, CA, USA. [email protected].
  • 10. Gladstone Institutes, University of California, San Francisco, San Francisco, CA, USA. [email protected].
  • 11. Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, CA, USA. [email protected].
  • 12. Department of Chemistry, University of California, Berkeley, Berkeley, CA, USA. [email protected].
  • 13. MBIB Division, Lawrence Berkeley National Laboratory, Berkeley, CA, USA. [email protected].
  • # Contributed equally.
PMID: 41034576 DOI: 10.1038/s41586-025-09569-9
Abstract

Microbial and viral co-evolution has created immunity mechanisms involving oligonucleotide signalling that share mechanistic features with human Antiviral systems1. In these pathways, including cyclic oligonucleotide-based antiphage signalling systems (CBASSs) and type III CRISPR systems in bacteria and cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) in humans, oligonucleotide synthesis occurs upon detection of virus or foreign genetic material in the cell, triggering the Antiviral response2-4. Here, in an unexpected inversion of this process, we show that the CRISPR-related enzyme mCpol synthesizes cyclic oligonucleotides constitutively as part of an active mechanism that represses a toxic effector. Cell-based experiments demonstrated that the absence or loss of mCpol-produced cyclic oligonucleotides triggers cell death, preventing the spread of viruses that attempt immune evasion by depleting host cyclic nucleotides. Structural and mechanistic investigation revealed mCpol to be a di-adenylate cyclase whose product, c-di-AMP, prevents toxic oligomerization of the effector protein 2TMβ. Analysis of cells by fluorescence microscopy showed that lack of mCpol allows 2TMβ-mediated cell death due to inner membrane collapse. These findings unveil a powerful defence strategy against virus-mediated immune suppression, expanding our understanding of the role of oligonucleotides in immunity.

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