A transdermal CRISPR/Cas9 strategy using biodegradable microneedles to target YB-1 in melanoma

Melanoma's aggressiveness and therapeutic resistance highlight the need for innovative strategies. This study developed a CRISPR/Cas9 ribonucleoprotein (RNP) system targeting the oncogenic YB-1 gene, combined with doxorubicin (DOX), delivered by chondroitin sulfate (CS)-based microneedles (MN) incorporating designed nanoparticles (CNPs). YB-1, a critical oncogene driving melanoma progression, metastasis, and drug resistance, was targeted to enhance therapy efficacy. In vitro experiments demonstrated that CNPs enhanced cellular uptake efficiency, achieving a dual-drug delivery rate of 74.4% and mediating 65.32% gene editing efficiency. The application of CNPs@DOX@RNP reduced tumor cell viability to just 3.51%, while simultaneously lowering off-target toxicity. In vivo C57BL/6 melanoma models showed significant tumor growth inhibition, increased apoptosis, and suppressed proliferation/angiogenesis. Following treatment with CNPs@DOX@RNP/MN, tumor weight exhibited a 92.56% reduction relative to the untreated control. Biocompatibility assays confirmed safety with minimal organ damage. This MN platform offers a synergistic, safe, and effective strategy for combined YB-1 gene editing and chemotherapy, addressing melanoma's therapeutic challenges.