A facile method was designed that can specifically deliver CRISPR/Cas9 into target cells nuclei and reduce the off-target effects. A multifunctional delivery vector for FOXM1 knockout was composed by integration of cell targeting polymer (hyaluronic acid) and cell and nuclear targeting group (AS1411 aptamer) on the surface of nanoparticles formed by genome editing plasmid and chitosan (CS) as the core (Apt-HA-CS-CRISPR/Cas9). The data of cytotoxicity experiment and western blot confirmed this issue. The results of flow cytometry analysis and fluorescence imaging demonstrated that Apt-HA-CS-CRISPR/Cas9 was significantly internalized into target cells (MCF-7, SK-MES-1, HeLa) but not into nontarget cells (HEK293). Furthermore, the in vivo studies displayed that the Apt-HA-CS-CRISPR/Cas9 was strongly rendered tumor inhibitory effect and delivered efficiently CRISPR/Cas9 into the tumor with no detectable distribution in other organs compared with naked plasmid. This approach provides an avenue for specific in vivo gene editing therapeutics with the lowest side effect.

设计了一种简便的方法，可以将 CRISPR/Cas9 特异性递送到靶细胞核中并减少脱靶效应。基因组编辑质粒、壳聚糖（CS）为核心（Apt- HA-CS-CRISPR/Cas9)。细胞毒性实验和western blot的数据证实了这个问题。流式细胞术分析和荧光成像结果表明，Apt-HA-CS-CRISPR/Cas9 显着内化到靶细胞（MCF-7、SK-MES-1、HeLa）中，但不内化到非靶细胞（HEK293）中。此外，体内研究表明，与裸质粒相比，Apt-HA-CS-CRISPR/Cas9 具有很强的肿瘤抑制作用，并将 CRISPR/Cas9 有效地递送到肿瘤中，在其他器官中没有可检测到的分布。这种方法为具有最低副作用的特定体内基因编辑疗法提供了途径。