Aptamer-mediated CRISPR-Cas9 delivery for precision oncology: integrating molecular targeting, intracellular trafficking, and nanocarrier engineering

Abstract

Cancer therapy remains constrained by insufficient molecular specificity, intratumoral heterogeneity, and the emergence of therapeutic resistance that limit durable clinical responses. CRISPR Cas9 genome editing offers an unprecedented opportunity to directly reprogram oncogenic drivers, tumor suppressor pathways, and regulatory noncoding elements with nucleotide level precision. Nevertheless, its clinical translation is fundamentally restricted by inefficient tumor selective delivery, limited intracellular trafficking, and concerns surrounding off target genome modification. Aptamers have emerged as highly adaptable targeting ligands capable of addressing these barriers through selective recognition of tumor associated surface receptors, minimal immunogenicity, and favorable physicochemical properties that support deep tissue penetration. Structure guided molecular docking has enabled rational optimization of aptamer receptor interactions, facilitating improved binding energetics and target discrimination prior to experimental implementation. In parallel, elucidation of receptor mediated endocytosis pathways and endosomal escape mechanisms has advanced understanding of how aptamer functionalization enhances intracellular transport and nuclear access of CRISPR components. Recent progress in nanotechnology has further expanded delivery strategies through lipid nanoparticles, polymeric and inorganic carriers, extracellular vesicles, and hybrid viral–nonviral platforms engineered with refined PEG architectures and linker chemistries to enhance stability and controlled release. Integration of these approaches enables spatiotemporally controlled genome editing within complex tumor microenvironments and mitigates biological obstacles such as tumor-associated extracellular matrix–mediated resistance and systemic degradation of gene editing cargos. This review highlights recent integrated advancements in aptamer engineering, structure-guided molecular docking, understanding of intracellular trafficking and endosomal escape mechanisms, and nanocarrier design. Collectively, these developments position aptamer-guided CRISPR-Cas9 systems as a promising strategy to improve tumor-specific delivery and genome editing precision in cancer therapy. However, substantial challenges remain regarding in vivo stability, manufacturing scalability, off-target effects, and rigorous preclinical and clinical validation before broad clinical translation can be realized.