A DNA/Upconversion Nanoparticle Complex Enables Controlled Co‐Delivery of CRISPR‐Cas9 and Photodynamic Agents for Synergistic Cancer Therapy

Abstract Photodynamic therapy (PDT) depends on the light‐irradiated exciting of photosensitizer (PS) to generate reactive oxygen species (ROS), which faces challenges and limitations in hypoxia and antioxidant response of cancer cells, and limited tissue‐penetration of light. Herein, a multifunctional DNA/upconversion nanoparticles (UCNPs) complex is developed which enables controlled co‐delivery of CRISPR‐Cas9, hemin, and protoporphyrin (PP) for synergistic PDT. An ultralong single‐stranded DNA (ssDNA) is prepared via rolling circle amplification (RCA), which contains recognition sequences of single guide RNA (sgRNA) for loading Cas9 ribonucleoprotein (RNP), G‐quadruplex sequences for loading hemin and PP, and linker sequences for combining UCNP. Cas9 RNP cleaves the antioxidant regulator nuclear factor E2‐related factor 2 (Nrf2), improving the sensitivity of cancer cells to ROS, and enhancing the synergistic PDT effect. The G‐quadruplex/hemin DNAzyme mimicks horseradish peroxidase (HRP) to catalyze the endogenous H 2 O 2 to O 2 , overcoming hypoxia condition in tumors. The introduced UCNP converts NIR irradiation with deep tissue penetration to light with shorter wavelength, exciting PP to transform the abundant O 2 to 1 O 2 . The integration of gene editing and PDT allows substantial accumulation of 1 O 2 in cancer cells for enhanced cell apoptosis, and this synergistic PDT has shown remarkable therapeutic efficacy in a breast cancer mouse model.