在这项研究中，通过将阳离子脂质 1,2-二油酰-3-三甲基铵-丙烷 (DOTAP) 与间充质干细胞膜 (MSCM) 相结合来构建一个平台，以产生带正电荷的混合囊泡。制备的混合囊泡用于浓缩用于生存素（BIRC5）基因编辑的BIRC5 CRISPR/Cas9质粒。然后通过静电相互作用将 Sgc8-c 适体（针对蛋白酪氨酸激酶 7）附着到制备的 NP 的表面。在这方面，可以靶向过度表达PTK7受体的黑色素瘤癌细胞（B16F0细胞系）。使用带有 B16F0 肿瘤的 C57BL/6 J 小鼠对该系统进行研究，以评估其转染效率、细胞毒性和临床前阶段的治疗性能。结果验证了Hybrid/BIRC5与Liposome/BIRC5相比在细胞毒性和转染效率方面的优越性。细胞暴露于 Hybrid/BIRC5 后细胞毒性显着增强。此外，用 Apt-Hybrid/BIRC5 处理的细胞对 PTK7 阳性 B16F0 癌细胞表现出比 PKT7 阴性 CHO 细胞系更高的抗增殖活性。蛋白质印迹分析证实，活性肿瘤靶向纳米颗粒通过下调 BIRC5 表达来增加细胞毒性。在临床前阶段，Apt-Hybrid/BIRC5 对 B16F0 肿瘤小鼠表现出显着的肿瘤生长抑制作用。因此，我们的研究表明，通过 CRISPR/Cas9 系统对 BIRC5 进行基因组编辑可以为黑色素瘤癌症治疗提供一种潜在安全的方法，并且具有巨大的临床转化潜力。版权所有 © 2024 Elsevier Inc. 保留所有权利。

In this study, a platform was fabricated by combining a cationic lipid, 1,2-Dioleoyl-3-trimethylammonium-propane (DOTAP) with mesenchymal stem cell membrane (MSCM) to produce a positively charged hybrid vesicle. The prepared hybrid vesicle was used to condense BIRC5 CRISPR/Cas9 plasmid for survivin (BIRC5) gene editing. The Sgc8-c aptamer (against protein tyrosine kinase 7) was then attached to the surface of the prepared NPs through electrostatic interactions. In this regard, melanoma cancer cells (B16F0 cell line) overexpressing PTK7 receptor could be targeted. Investigations were conducted on this system to evaluate its transfection efficiency, cellular toxicity, and therapeutic performance in preclinical stage using B16F0 tumor bearing C57BL/6 J mice. The results verified the superiority of the Hybrid/ BIRC5 compared to Liposome/ BIRC5 in terms of cellular toxicity and transfection efficiency. The cells exposure to Hybrid/BIRC5 significantly enhanced cytotoxicity. Moreover, cells treated with Apt-Hybrid/BIRC5 showed higher anti-proliferation activity toward PTK7-positive B16F0 cancer cells than that of the PKT7-negative CHO cell line. The active tumor targeting nanoparticles increased the cytotoxicity through down-regulation of BIRC5 expression as confirmed by Western blot analysis. In preclinical stage, Apt-Hybrid/BIRC5 showed remarkable tumor growth suppression toward B16F0 tumorized mice. Thus, our study suggested that genome editing for BIRC5 through the CRISPR/Cas9 system could provide a potentially safe approach for melanoma cancer therapy and has great potential for clinical translation.Copyright © 2024 Elsevier Inc. All rights reserved.