Short-pulse laser pretreatment induced transduction of adeno-associated viral (AAV) vector into retinal pigment epithelial (RPE) cells. Shown are representative confocal microscopic RPE wholemount images (200×) of the control (laser-untreated/vector-uninjected, laser-treated/vector-uninjected, and laser-untreated/vector-injected; A–E) and scAAV type 2-, 5-, and 8-treated eyes after laser pretreatment (F–H). Morphological changes of RPE cells were also observed at the lasered region. AAV5 vector (G) transduced fewer RPE cells compared with AAV2 (F) and AAV8 (H). Several RPE cells adjacent to the laser site were also transduced by AAV vectors. Laser-untreated/vector-uninjected and laser-treated/vector-uninjected control eyes did not show any EGFP expression (A and E), while laser-untreated/vector-injected eyes demonstrated few of sporadic EGFP expression in the RPE cells. The wholemount was processed with β-catenin (red) immunostaining to show the intact cell-to-cell junction. Some of the RPE cells showed cytoplasmic accumulation of β-catenin, indicating the activation of canonical Wnt signaling pathway. *Center of laser-treated region. The scale bars represent 100 μm.