Recombinant adeno-associated virus (AAV) vectors predominantly remain episomal and integrate randomly at low frequency, raising safety concerns for gene therapy. We developed an rDNA-targeted AAV vector designed for site-specific integration via homologous recombination. An hFah expression cassette flanked by rDNA homology rescued Fah⁻/⁻ mice at significantly lower doses than conventional AAV, with stable phenotypic correction confirmed by serial transplantation and site-specific junction PCR. Targeted vectors showed enhanced persistence after partial hepatectomy and enabled systemic site-specific integration in multiple tissues. This strategy provides a highly efficient, low-dose platform for durable gene therapy.
