Deep parallel characterization of AAV tropism and AAV-mediated transcriptional changes via single-cell RNA sequencing

Transcriptomic analysis revealed that this neuronal bias is due mainly to increased targeting efficiency for glutamatergic neurons, which we confirmed by RNA fluorescence in situ hybridization. We further uncovered cell subtype tropisms of AAV variants in vascular and glial cells, such as low transduction of pericytes and Myoc+ astrocytes. Additionally, we have observed cell-type-specific transitory responses to systemic AAV-PHP.eB administration, such as upregulation of genes involved in p53 signaling in endothelial cells three days post-injection, which return to control levels by day twenty-five.

The presented experimental and computational approaches for parallel characterization of AAV tropism with ability to simultaneously measure the transcriptional response of transduction will facilitate the advancement of safe and precise gene delivery vehicles.