Bioorthogonal Reaction‐Mediated Tumor‐Selective Delivery of CRISPR/Cas9 System for Dual‐Targeted Cancer Immunotherapy

Abstract CRISPR system‐assisted immunotherapy is an attractive option in cancer therapy. However, its efficacy is still less than expected due to the limitations in delivering the CRISPR system to target cancer cells. Here, we report a new CRISPR/Cas9 tumor‐targeting delivery strategy based on bioorthogonal reactions for dual‐targeted cancer immunotherapy. First, selective in vivo metabolic labeling of cancer and activation of the cGAS‐STING pathway was achieved simultaneously through tumor microenvironment (TME)‐biodegradable hollow manganese dioxide (H‐MnO 2 ) nano‐platform. Subsequently, CRISPR/Cas9 system‐loaded liposome was accumulated within the modified tumor tissue through in vivo click chemistry, resulting in the loss of protein tyrosine phosphatase N2 (PTPN2) and further sensitizing tumors to immunotherapy. Overall, our strategy provides a modular platform for precise gene editing in vivo and exhibits potent antitumor response by boosting innate and adaptive antitumor immunity.