Mannose-functionalized star polycation mediated CRISPR/Cas9 delivery for lung cancer therapy

Abstract

The survivin gene, highly expressed in most cancer cells, is closely associated with inhibiting apoptosis. Therefore, gene editing for the survivin gene has great potential in tumor therapy. However, it is difficult for plasmid DNA (pDNA) to be taken up directly by cells, and thus the construction of gene vectors is a key step for successful gene editing. Ethanolamine-functionalized polyglycidyl methacrylate (PGEA) has been proved to facilitate the transfection of pDNA into cells in both in vivo and in vitro experiments. However, PGEA does not specifically recognize tumor cells. Some tumor cells express more mannose receptor (MR) than healthy cells. To achieve efficient target and transfection, we designed mannose-functionalized four-arm PGEA cationic polymers (P(GEA-co-ManMA), GM) with different molecular weights. GM was combined with pCas9-survivin. The mannose unit of GM/pCas9-survivin was identified by MR to selectively enter lung cancer cells. In vitro experiments showed that GM not only had excellent biocompatibility, gene transfection performance, and targeted ability, but also significantly inhibited the proliferation of tumor cells when used in combination with pCas9-survivin. At the same time, we also studied the relationship between the molecular weight and therapeutic effect.