Background Epiretinal membranes in patients with proliferative vitreoretinopathy (PVR) consist of extracellular matrix and a number of cell types including retinal pigment epithelial (RPE) cells and fibroblasts, whose contraction causes retinal detachment. In RPE cells depletion of platelet-derived growth factor (PDGF) receptor (PDGFR) beta suppresses vitreous-induced Akt activation, whereas in fibroblasts Akt activation through indirect activation of PDGFR alpha by growth factors outside the PDGF family (non-PDGFs) plays an essential role in experimental PVR. Whether non-PDGFs in the vitreous, however, were also able to activate PDGFR beta in RPE cells remained elusive. Methods The CRISPR/Cas9 technology was utilized to edit a genomic PDGFRB locus in RPE cells derived from an epiretinal membrane (RPEM) from a patient with PVR, and a retroviral vector was used to express a truncated PDGFR beta short of a PDGF-binding domain in the RPEM cells lacking PDGFR beta. Western blot was employed to analyze expression of PDGFR beta and alpha-smooth muscle actin, and signaling events (p-PDGFR beta and p-Akt). Cellular assays (proliferation, migration and contraction) were also applied in this study. Results Expression of a truncated PDGFR beta lacking a PDGF-binding domain in the RPEM cells whose PDGFRB gene has been silent using the CRISPR/Cas9 technology restores vitreous-induced Akt activation as well as cell proliferation, epithelial-mesenchymal transition, migration and contraction. In addition, we show that scavenging reactive oxygen species (ROS) with N-acetyl-cysteine and inhibiting Src family kinases (SFKs) with their specific inhibitor SU6656 blunt the vitreous-induced activation of the truncated PDGFR beta and Akt as well as the cellular events related to the PVR pathogenesis. These discoveries suggest that in RPE cells PDGFR beta can be activated indirectly by non-PDGFs in the vitreous via an intracellular pathway of ROS/SFKs to facilitate the development of PVR, thereby providing novel opportunities for PVR therapeutics. Conclusion The data shown here will improve our understanding of the mechanism by which PDGFR beta can be activated by non-PDGFs in the vitreous via an intracellular route of ROS/SFKs and provide a conceptual foundation for preventing PVR by inhibiting PDGFR beta transactivation (ligand-independent activation).