Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) technology enables targeted gene editing, but cancer gene therapy with this approach requires improvements to enable safe and efficient delivery of CRISPR/Cas9 to tumors. We developed and evaluated a self-assembled liposome to selectively deliver CRISPR/Cas9 to cancer tissues. Our CRISPR/Cas9 system effectively inhibited proliferation of human papillomavirus (HPV) 16-positive cervical cancer cells and induced apoptosis by inactivating the HR-HPV16E6/E7 oncogene. Based on this system, we prepared a long-circulating pH-sensitive cationic nano-liposome complex with a high cell targeting and gene knockout rate. Intratumoral injection of cationic liposomes targeted to splicing HPV16 E6/E7 in nude mice significantly inhibited tumor growth without significant toxicity in vivo. Liposomes that targeted HPV16 E6/E7 splicing were established as a basis for treatment of HPV16-positive cervical cancer drug candidates. Our study demonstrates that this liposome offers an efficient delivery system for nonviral gene editing, adding to the armamentarium of gene editing tools to advance safe and effective precision medicine-based cancer therapeutics.

中文翻译：

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使用由 pH 敏感的阳离子脂质体提供的成簇规则散布短回文重复序列/Cas9 对人乳头瘤病毒致癌基因进行操作。

成簇的规则间隔短回文重复序列 (CRISPR)/CRISPR 相关蛋白 9 (Cas9) 技术可实现靶向基因编辑，但使用这种方法的癌症基因治疗需要改进，才能安全有效地将 CRISPR/Cas9 递送至肿瘤。我们开发并评估了一种自组装脂质体，以选择性地将 CRISPR/Cas9 递送至癌症组织。我们的 CRISPR/Cas9 系统通过灭活 HR-HPV16E6/E7 癌基因，有效抑制人乳头瘤病毒 (HPV) 16 阳性宫颈癌细胞的增殖并诱导细胞凋亡。基于该系统，我们制备了具有高细胞靶向性和基因敲除率的长循环pH敏感性阳离子纳米脂质体复合物。在裸鼠中肿瘤内注射靶向剪接 HPV16 E6/E7 的阳离子脂质体可显着抑制肿瘤生长，而在体内无显着毒性。靶向 HPV16 E6/E7 剪接的脂质体被确立为治疗 HPV16 阳性宫颈癌候选药物的基础。我们的研究表明，这种脂质体为非病毒基因编辑提供了一个有效的传递系统，增加了基因编辑工具的军备，以推进安全有效的基于精准医学的癌症治疗。