Gene therapy has drawn great attention in the treatments of many diseases, especially for cardiovascular diseases. However, the development of gene carriers with low cytotoxicity and multitargeting function is still a challenge. Herein, the multitargeting REDV-G-TAT-GNLS peptide was conjugated to amphiphilic cationic copolymer poly(ε-caprolactone-co-3(S)-methyl-morpholine-2,5-dione)-g-polyethyleneimine (PCLMD-g-PEI) via a heterobifunctional orthopyridyl disulfide-poly(ethylene glycol)-N-hydroxysuccinimide (OPSS-PEG-NHS) linker to prepare PCLMD-g-PEI-PEG-REDV-G-TAT-G-NLS copolymers with the aim to develop the gene carriers with low cytotoxicity and high transfection efficiency. The multitargeting micelles were prepared from PCLMD-g-PEI-PEG-REDV-G-TAT-G-NLS copolymers by selfassembly method and used to load pEGFP-ZNF580 plasmids (pDNA) to form gene complexes for enhancing the proliferation and migration of endothelial cells (ECs). The loading pDNA capacity was proved by agarose gel electrophoresis assay. These multitargeting gene complexes exhibited low cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The high internalization efficiency of these gene complexes was confirmed by flow cytometry. The results of in vitro transfection demonstrated that these multitargeting gene complexes possessed relatively high transfection efficiency. The rapid migration of ECs transfected by these gene complexes was verified by wound healing assay. Owing to ECs-targeting ability, cell-penetrating ability and nuclear targeting capacity of REDV-G-TAT-G-NLS peptide, the multitargeting polycationic gene carrier with low cytotoxicity and high transfection efficiency has great potential in gene therapy.

基因疗法在许多疾病，尤其是心血管疾病的治疗中引起了极大的关注。然而，具有低细胞毒性和多靶功能的基因载体的开发仍然是一个挑战。这里，multitargeting REDV-G-TAT-GNLS肽缀合于两亲性阳离子共聚物聚（ε -己内酯-共-3-（S） -甲基吗啉-2,5-二酮） -克-聚乙烯亚胺（PCLMD-克-通过异双功能邻吡啶基二硫化物-聚（乙二醇）-N-羟基琥珀酰亚胺（OPSS-PEG-NHS）接头制备PCLMD- g -PEI-PEG-REDV-G-TAT-G-NLS共聚物具有低细胞毒性和高转染效率的基因载体。从PCLMD- g制备多目标胶束-PEI-PEG-REDV-G-TAT-G-NLS共聚物采用自组装方法，用于加载pEGFP-ZNF580质粒（pDNA）形成基因复合物，以增强内皮细胞（EC）的增殖和迁移。通过琼脂糖凝胶电泳分析证明了pDNA的负载能力。这些多靶基因复合物通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基溴化四氮唑测定显示出低细胞毒性。通过流式细胞术证实了这些基因复合物的高内在化效率。体外转染的结果表明，这些多靶基因复合物具有较高的转染效率。通过伤口愈合试验证实了被这些基因复合物转染的ECs的快速迁移。由于具有EC定位能力，