MicroRNA-responsive release of Cas9/sgRNA from DNA nanoflower for cytosolic protein delivery and enhanced genome editing
MicroRNA-responsive release of Cas9/sgRNA from DNA nanoflower for cytosolic protein delivery and enhanced genome editing
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DNA 纳米花中 MicroRNA 响应性释放 Cas9/sgRNA，用于胞浆蛋白递送和增强基因组编辑
DOI：
10.1016/j.biomaterials.2020.120221
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发表时间：
2020-10-01
期刊：
影响因子：
14
通讯作者：
Zhang, Kaixiang
中科院分区：
文献类型：
--
作者：
Shi, Jinjin;Yang, Xue;Zhang, Kaixiang
基于纳米颗粒的 CRISPR/Cas9 递送系统为遗传性疾病的特异性和精确治疗带来了巨大的希望。在此，我们开发了一种基于 DNA 纳米花的平台，用于将 Cas9/sgRNA 复合物响应 microRNA 胞质递送到肿瘤细胞中。生物相容性DNA纳米载体可以通过序列杂交有效负载Cas9/sgRNA。重要的是，这种杂交可以通过立足点介导的链置换过程被肿瘤特异性 miRNA 取代，并实现 DNA 纳米花中 Cas9/sgRNA 的细胞类型特异性释放。我们已经证实，与非响应性对照相比，这种 miRNA 响应性释放过程可以显着提高基因组编辑效率。该策略提出了一种通过结合刺激响应性 Cas9/sgRNA 释放过程来设计更特异、更高效的基于 CRISPR 的基因组治疗系统的通用方法。
Nanoparticle-based CRISPR/Cas9 delivery systems hold great promise for specific and precise treatment of genetic disorder diseases. Herein, we developed a DNA nanoflower-based platform for microRNA-responsive cytosolic delivery of Cas9/sgRNA complex into tumor cells. The biocompatible DNA nano-vehicles can efficiently load Cas9/sgRNA by sequence hybridization. Importantly, this hybridization can be replaced by a tumor specific miRNA through toehold-mediated strand displacement process and achieve cell-type-specific release of Cas9/sgRNA from the DNA nanoflowers. We have verified that this miRNA-responsive releasing process can significantly improve the genome editing efficiency comparing with non-responsive control. This strategy suggests a versatile way for designing more specific and efficient CRISPR-based genome therapy system by incorporating stimuli-responsive Cas9/sgRNA release process.