CRISPR/Cas9 is a promising technology for gene editing. To date, intracellular delivery vehicles for CRISPR/Cas9 are limited by issues of immunogenicity, restricted packaging capacity, and low tolerance. Here, we report an alternative, nonviral delivery system for CRISPR/Cas9 based on engineered exosomes. We show that non-autologous exosomes can encapsulate CRISPR/Cas9 plasmid DNA via commonly available transfection reagents and can be delivered to recipient cancer cells to induce targeted gene deletion. As a proof-of-principle, we demonstrate that exosomes loaded with CRISPR/Cas9 can target the mutant Kras ^G12D oncogenic allele in pancreatic cancer cells to suppress proliferation and inhibit tumor growth in syngeneic subcutaneous and orthotopic models of pancreatic cancer. Exosomes may thus be a promising delivery platform for CRISPR/Cas9 gene editing for targeted therapies.

中文翻译：

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外泌体介导的 CRISPR/Cas9 递送用于靶向胰腺癌中的致癌 KrasG12D。

CRISPR/Cas9 是一种很有前途的基因编辑技术。迄今为止，CRISPR/Cas9 的细胞内递送载体受到免疫原性、包装容量受限和耐受性低等问题的限制。在这里，我们报告了一种基于工程外泌体的 CRISPR/Cas9 替代非病毒传递系统。我们表明，非自体外泌体可以通过常用的转染试剂包裹 CRISPR/Cas9 质粒 DNA，并可以递送至受体癌细胞以诱导靶向基因缺失。作为原理验证，我们证明装载 CRISPR/Cas9 的外泌体可以靶向突变体Kras ^G12D胰腺癌细胞中的致癌等位基因抑制胰腺癌同系皮下和原位模型中的增殖和抑制肿瘤生长。因此，外泌体可能是用于靶向治疗的 CRISPR/Cas9 基因编辑的有前途的递送平台。