Herein, five aggregation-induced emission (AIE) photosensitizers (PSs) with D-π-A structures are smoothly designed and synthesized through donor and acceptor engineering. The photophysical properties and theoretical calculation results show that the synergistic effect of methoxy substituted tetraphenylethene (MTPE), 3,4-ethylenedioxythiophene can enhance the intramolecular charge transfer effect (ICT), and promote the intersystem crossing (ISC) process of the whole molecule. In these AIE-PSs, the best-performing AIE-PS (MTPE-DT-Py) has bright NIR (740 nm) emission, the highest 1O2 generation efficiency (5.9-fold that of Rose Bengal) and efficient mitochondrial targeting ability. Subsequently, PDT anti-cancer and anti-bacterial experiments indicate that MTPE-DT-Py could obviously target mitochondria and kill breast cancer cells (MCF-7), and selectively inactivate S. aureus (G(+)) under white light irradiation. This work mainly proposes a practical design strategy for high effect AIE-PSs and provides more excellent candidates for fluorescence imaging-guided photodynamic therapy.