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bioRxiv · 10.64898/2026.10.26.761871

Single-vector AAV delivery of En-ISPmu1 and donor DNA for in vivo targeted genome integration

Conci, A.·Bonuzzi, I.·Ciciani, M.·Esposito, F.·Piccolo, P.·Collaud, F.·Ronzitti, G.·Auricchio, A.·Segata, N.·Cereseto, A.
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Abstract

TnpBs are compact RNA-guided nucleases ideally suited for in vivo delivery. By screening novel TnpBs identified from large metagenomic datasets, ISPmu1 was discovered and its protein and {omega}RNA were engineered to generate En-ISPmu1, which achieved editing efficiencies exceeding 65% across 12 human loci. Owing to its compact size, En-ISPmu1 was delivered together with a donor DNA in a single AAV vector, yielding up to 12.7% targeted integration in mouse liver. These findings establish En-ISPmu1 as a promising versatile platform for in vivo therapeutic genome editing applications.

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Conci, A., Bonuzzi, I., Ciciani, M., Esposito, F., Piccolo, P., Collaud, F., Ronzitti, G., Auricchio, A., Segata, N., Cereseto, A.. 2026-10-28. Single-vector AAV delivery of En-ISPmu1 and donor DNA for in vivo targeted genome integration. https://doi.org/10.64898/2026.10.26.761871

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