Tumor targeted genome editing mediated by a multi-functional gene vector for regulating cell behaviors.

For effective regulation of cell behaviors and prevention of tumor development by genome editing, we constructed multi-functional self-assembled nanoparticles based on natural polymers to deliver CRISPR-Cas9 plasmid to tumorous cells. The CRISPR based gene editing plasmid to knockout CDK11 gene was complexed with protamine sulfate, and then the complex was decorated by a multi-functional outer layer composed of an endosomolytic peptide (KALA) and aptamer AS1411 incorporated carboxymethyl chitosan. The resultant multi-functional nanoparticles, which exhibit significantly enhanced delivery efficiency, can specifically deliver the plasmid into tumor cell nuclei owing to the favorable effects of KALA in cellular uptake and endosomal escape, together with the cancer cell and cell nucleus targeting capability of AS1411 ligands. The genome editing mediated by the nanoparticles leads to a dramatic decrease (>75%) in CDK11 expression, which results in further modulation of cancer cells with significant down-regulation of the proteins (MMP-9 and VEGF) involved in tumor development and metastasis as well as up-regulation of the tumor suppressor protein p53. More importantly, the detection of immune-related proteins after genome editing shows that the significantly enhanced Fas, CD80, MICA, MICB, and HLA-1 expression and decreased CD47 and MUC1 expression, indicating the genome editing is favorable for reversal of tumor-induced immunosuppression and prevention of tumor development.