NOVEL VIRUS-LIKE PARTICLES FOR NUCLEAR DNA DELIVERY

基因治疗是许多获得性和遗传性疾病最有希望的治疗方法。由于DNA递送效率不足，其全部潜力尚未实现。非病毒载体是合成载体，其不存在由病毒引起的大多数问题，但也缺乏其许多期望的性质（有效的细胞摄取、内体释放、定向细胞质移动、基因组的拆卸和核递送）。如果人工基因传递粒子能够模仿这些优点，那将大大提高它们的性能。在拟议的第一阶段研究期间，将合成新型阳离子聚合物并用于形成各种凝聚的DNA颗粒。然后，将使用Mirus先前开发的两种技术对颗粒进行修饰：对表面进行充电以获得负zeta电位，但保持完全凝聚的DNA，并将颗粒置于笼中以防止其聚集。还将创建具有近中性表面的粒子。将在生理缓冲液和胞质提取物以及显微注射细胞的细胞质中评估其稳定性。拟定商业应用：在拟议的研究中开发的核转运技术将被纳入含有细胞受体和内体溶解剂的配体的基因治疗载体中。 鉴于非病毒载体工业认识到核运输障碍的重要性，克服这一障碍的技术将具有巨大的商业价值。

Gene therapy is the most promising treatment for many acquired and genetic diseases. Its full potential has not been realized due to the inadequate efficiency of DNA delivery. Non-viral vectors are synthetic vehicles which are void of most problems caused by viruses, but also lack many of their desirable properties (efficient cellular uptake, endosomal release, directional cytoplasmic movement, disassembly and nuclear delivery of the genome). If artificial gene delivery particles could mimic such virtues, those would greatly improve their performance. During the proposed Phase I studies novel cationic polymers will be synthesized and used for the formation of a variety of condensed DNA particles. The particles will then be modified by using two technologies previously developed by Mirus: recharging the surface to obtain negative zeta potential, yet maintaining fully condensed DNA, and caging the particles to prevent their aggregation. Particles with near-neutral surface will also be created. Their Stability will be assessed in physiological buffers and cytosolic extracts, and also in the cytoplasm of microinjected cells. PROPOSED COMMERCIAL APPLICATION: The nuclear transport technology developed in the proposed studies will be incorporated into gene therapy vectors that contain ligands for cellular receptors and endosomolytic agents. Given that the non-viral vector industry realizes the importance of the nuclear transport hurdle, technology for overcoming this hurdle will have great commercial value.