Light‐Guided Genetic Scissors Based on Phosphorene Quantum Dot

Abstract Genetic engineering faces persistent challenges in achieving precise Deoxyribonucleic acid (DNA) cleavage, especially with the limitations associated with current enzyme‐based methods, exemplified by issues in CRISPR technologies. This study introduces a groundbreaking approach: utilizing reactive oxygen species (ROS) generated by Multiphoton Absorption (MPA)‐excited Black Phosphorus Quantum Dots (BPQDs) under femto‐second laser irradiation. This innovative method not only allows excitation with lower energy but also enhances overall efficiency. The integration of complementary RNA sequences facilitates high‐efficiency, site‐selective DNA cleavage in the system, named “TADPOLE” (Targeted DNA Precision Oriented Laser Excision). Beyond its precision in targeting arbitrary DNA sequences using quantum dots, TADPOLE harnesses the unique multiphoton absorption property of BPQDs, enabling excitation with lower‐energy light sources suitable for in vivo applications in the future. Moreover, the approach integrates guiding RNA and ultrafast laser technology to provide precise control over local ROS generation and minimal heat production. This guarantees site‐specific DNA cleavage while mitigating the risk of damage to non‐targeted sequences. In summary, this study catalyzes advancements in enzyme‐free DNA cleavage technologies, with transformative implications for genetic engineering, biotechnology, and medicine. The holistic precision, versatility, and endurance presented by TADPOLE open new avenues for targeted gene therapies and transformative applications in related fields.