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前途的治疗方法。由于DNA递送效率不高，它的全部潜力尚未实现。非病毒载体是一种合成载体，它不存在病毒引起的大多数问题，但也缺乏许多理想的特性（有效的细胞摄取、内体释放、细胞质定向运动、基因组的拆卸和核递送）。如果人工基因传递颗粒能够模仿这些优点，将大大提高它们的性能。在拟议的第一阶段研究中，将合成新型阳离子聚合物并用于形成各种浓缩DNA颗粒。然后，这些颗粒将通过Mirus先前开发的两种技术进行修饰：给表面充电以获得负的zeta电位，同时保持完全浓缩的DNA，并将颗粒关在笼子里以防止它们聚集。具有接近中性表面的粒子也将被创造出来。它们的稳定性将在生理缓冲液和细胞质提取物中进行评估，也将在微注射细胞的细胞质中进行评估。拟议的商业应用：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配体和内溶剂的基因治疗载体中。鉴于非病毒载体行业意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.