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

En-ISPmu1 enables genome editing efficiencies exceeding 65% across 12 human loci and supports single-vector AAV delivery with donor DNA.

• Why it matters: In vivo therapeutic genome editing needs compact, RNA-guided nucleases that can be delivered efficiently while achieving targeted integration.
• What they did: Novel TnpBs from large metagenomic datasets were screened to discover ISPmu1, then engineered into En-ISPmu1 with its protein and omegaRNA; it was delivered with donor DNA in one AAV vector.
• The result: In mouse liver, single-vector delivery achieved up to 12.7% targeted integration, supporting En-ISPmu1 as a versatile in vivo genome editing platform.