Cas12a2 elicits abortive infection via RNA-triggered destruction of double-stranded DNA

Dmytrenko, O.; Neumann, G.; Hallmark, T.; Keiser, D. J.; Crowley, V. M.; Vialetto, E.; Mougiakos, I.; Wandera, K. G.; Domgaard, H.; Weber, J.; Metcalf, J.; Begemann, M. B.; Gray, B. N.; Jackson, R. N.; Beisel, C. L.

doi:10.1101/2022.06.13.495973

bioRxiv · 10.1101/2022.06.13.495973

Cas12a2 elicits abortive infection via RNA-triggered destruction of double-stranded DNA

Dmytrenko, O.·Neumann, G.·Hallmark, T.·Keiser, D. J.·Crowley, V. M.·Vialetto, E.·Mougiakos, I.·Wandera, K. G.·Domgaard, H.·Weber, J.·Metcalf, J.·Begemann, M. B.·Gray, B. N.·Jackson, R. N.·Beisel, C. L.

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Abstract

Bacterial abortive infection systems limit the spread of foreign invaders by shutting down or killing infected cells before the invaders can replicate1, 2. Several RNA-targeting CRISPR-Cas systems (e.g., types III and VI) cause Abi phenotypes by activating indiscriminate RNases3-5. However, a CRISPR-mediated abortive mechanism that relies on indiscriminate DNase activity has yet to be observed. Here we report that RNA targeting by the type V Cas12a2 nuclease drives abortive infection through non-specific cleavage of double-stranded (ds)DNA. Upon recognition of an RNA target with an activating protospacer-flanking sequence, Cas12a2 efficiently degrades single-stranded (ss)RNA, ssDNA, and dsDNA. Within cells, the dsDNase activity induces an SOS response and impairs growth, stemming the infection. Finally, we harnessed the collateral activity of Cas12a2 for direct RNA detection, demonstrating that Cas12a2 can be repurposed as an RNA-guided, RNA-targeting tool. These findings expand the known defensive capabilities of CRISPR-Cas systems and create additional opportunities for CRISPR technologies.

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Dmytrenko, O., Neumann, G., Hallmark, T., Keiser, D. J., Crowley, V. M., Vialetto, E., Mougiakos, I., Wandera, K. G., Domgaard, H., Weber, J., Metcalf, J., Begemann, M. B., Gray, B. N., Jackson, R. N., Beisel, C. L.. 2022-06-13. Cas12a2 elicits abortive infection via RNA-triggered destruction of double-stranded DNA. https://doi.org/10.1101/2022.06.13.495973

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