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RNA-guided nucleases enable a gene drive of insertion sequences in plasmids

Mears, K.; Rossine, F.; Quinones-Olvera, N.; Souque, C.; Baym, M.

2025-02-21 microbiology
10.1101/2025.02.20.638934 bioRxiv
Show abstract

Mobile genetic elements (MGEs) are diverse, self-replicating DNA molecules that can reside within cellular hosts and integrate into one another. This co-occurrence imposes distinct evolutionary pressures. Plasmids often contain insertion sequences (ISs). However, the multi-copy nature of plasmids should hinder IS introduction and spread, disfavoring inheritance of nascent plasmid variants through genetic drift. Mechanisms by which ISs overcome these barriers remain unidentified. Here we find that the RNA-guided nuclease TnpB enables such a mechanism to bias its inheritance in plasmids. We show that TnpB, the likely ancestor to Cas12, enables a gene drive to spread the IS within multicopy plasmids and functions as a primitive anti-self defense system in conjugative plasmids. The gene drive between TnpB-bearing ISs and plasmids promotes the spread of both MGEs beyond the ability of either individually. The nested existence between MGEs is not an incidental result of selfish spread, but a driver of it.

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