The emerging CRISPR/Cas9 system represents a promising platform for genome editing. However, its low transfection efficiency is a major problem hampering the application of the gene-editing potential of CRISPR/Cas9. Herein, by screening a pool of more than 56 kinds of agents, we constructed a novel polyethylene glycol phospholipid-modified cationic lipid nanoparticle (PLNP)-based delivery system that can condense and encapsulate a Cas9/single-guide RNA (sgRNA) plasmid (DNA) to form a core–shell structure (PLNP/DNA) that mediated up to 47.4% successful transfection of Cas9/sgPLK-1 plasmids in A375 cells in vitro. An intratumor injection of Cas9/sgPLK-1 plasmids into melanoma tumor-bearing mice resulted in significant downregulation of Polo-like kinase 1 (PLK-1) protein and suppression of the tumor growth (>67%) in vivo. This approach provides a versatile method that could be used for delivering the CRISPR/Cas9 system with high efficiency and safety both in vitro and in vivo.

新兴的CRISPR / Cas9系统代表了一个有前途的基因组编辑平台。然而，其低转染效率是阻碍CRISPR / Cas9基因编辑潜力的应用的主要问题。在此，我们通过筛选超过56种试剂的库，构建了一种新型的基于聚乙二醇磷脂修饰的阳离子脂质纳米颗粒（PLNP）的递送系统，该系统可以浓缩和封装Cas9 /单向导RNA（sgRNA）质粒（ DNA）形成核-壳结构（PLNP / DNA），可在体外介导A375细胞中成功将Cas9 / sgPLK-1质粒成功转染达47.4％。的瘤内注射Cas9 / sgPLK-1质粒的成黑色素瘤荷瘤小鼠导致的显著下调Polo样激酶1（PLK-1）的肿瘤生长的蛋白质和抑制（> 67％）在体内。这种方法提供了可用于具有高效率和安全性都递送CRISPR / Cas9系统的通用方法在体外和体内。