Focused ultrasound-mediated lipid nanoparticle delivery for brain gene editing

Efficient brain gene editing remains constrained by the lack of delivery platforms that combine efficacy, spatial precision, and translational potential. Compared with viral vectors, lipid nanoparticles (LNPs) offer larger cargo capacity and lower immunogenicity for repeat dosing. However, their brain delivery is restricted by the blood-brain barrier (BBB). Here, we show that focused ultrasound (FUS)-mediated BBB opening enables systemic delivery of CRISPR-encoding plasmid DNA (pDNA)-LNPs for brain gene editing. Using a pDNA construct containing astrocyte-targeting GfaABC1D promoter and dual guide RNAs targeting apolipoprotein E4 (APOE4), the strongest genetic risk factor for Alzheimer's disease, we achieved efficient APOE4 knockdown, with reduced APOE4 mRNA and apoE4 protein expression, and attenuated astrocytes and microglial activation. These results establish FUS-mediated pDNA-LNP delivery as a non-invasive, non-viral strategy for brain gene editing that provides spatial control and cell-type-specific expression, while accommodating large genetic payload and enabling repeatable dosing.