DNA CONDENSATION FOR IMPROVED STABILITY AND EXPRESSION

DNA 浓缩以提高稳定性和表达

• 批准号: 6076260
• 负责人: FRANK TAGLIAFERRI
• 金额: $ 10.27万
• 依托单位: VALENTIS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6076260
• 关键词: DNA damage; evaporation; gene expression; gene targeting; liposomes; plasmids; reporter genes; technology /technique development; tissue /cell culture; transfection

Non-viral gene delivery is an attractive alternative to viral delivery systems in that there have been relatively few reports of adverse effects associated with administration of non-viral delivery vehicles for DNA. Unfortunately, non-viral delivery systems (specifically cationic liposomes) have been hampered by inefficient gene delivery and inactivation by serum proteins upon intravenous administration. Preliminary data has shown that much of the low level expression can be attributed to the inefficient packaging of plasmid DNA within the delivery vehicle. Condensation of DNA with the cationic protein, protamine sulfate mimics the packaging of genetic material in mammals and produces a delivery system resembling an artificial sperm. This efficient condensation of DNA has been shown to result in an increase in the delivery of plasmid DNA 50-fold and provides protection from degradation by enzymes and nucleases. The ultimate goal of this research is to identify several condensation agents which will effectively package and protect DNA, allowing it to be incorporated into a cationic liposomal delivery system such that it may be safely and efficiently administered via a systemic route of injection. It is thought that this condensation process will impart stability and increased transgene activity necessary to overcome the inefficiencies of conventional non-viral gene delivery systems. PROPOSED COMMERCIAL APPLICATIONS: The research project proposed here will generate a greatly improved nonviral, liposomal gene delivery system which will be ubiquitously applicable to deliver genes, gene fragments, and oligonucleotides by various routes of administration including the parenteral and inhalatory route. It will, therefore, be applicable for gene therapy protocols of pulmonary diseases, metabolic diseases, cancer, and other diseases for which no therapeutic regimens are currently available.

非病毒基因递送是病毒递送系统的一种有吸引力的替代方案，因为与施用用于DNA的非病毒递送载体相关的不良反应的报道相对较少。不幸的是，非病毒递送系统（特别是阳离子脂质体）已经受到低效基因递送和静脉内施用时血清蛋白失活的阻碍。初步数据表明，许多低水平表达可归因于递送载体内质粒DNA的低效包装。硫酸鱼精蛋白是DNA与阳离子蛋白质的缩合物，它模仿哺乳动物遗传物质的包装，产生一种类似于人造精子的输送系统。这种DNA的有效缩合已显示导致质粒DNA的递送增加50倍，并提供保护以免受酶和核酸酶的降解。本研究的最终目标是确定几种缩合剂，这些缩合剂将有效地包装和保护DNA，使其能够掺入阳离子脂质体递送系统中，从而可以通过全身注射途径安全有效地给药。据认为，该缩合过程将赋予克服常规非病毒基因递送系统的低效性所必需的稳定性和增加的转基因活性。拟议的商业应用：本文提出的研究项目将产生一种大大改进的非病毒脂质体基因递送系统，该系统将普遍适用于通过各种给药途径（包括胃肠外和吸入途径）递送基因、基因片段和寡核苷酸。因此，它将适用于肺部疾病、代谢疾病、癌症和目前没有治疗方案的其他疾病的基因治疗方案。