IMPROVED NON VIRAL GENE TRANSFER VECTOR

描述(改编自调查人员摘要)：申请者有 提出了两个具有明显商业潜力的新观察结果。这个 首先观察到的是一项出人意料的发现，甲基化的质粒DNA 延长转基因在哺乳动物肺中表达的时间 细胞。这种表达的延伸很可能是由于保留了 转基因在细胞核中持续的时间长于 非甲基化质粒DNA。第二个观察结果是 在肺细胞中转a-1抗胰蛋白酶基因具有治疗作用 在性质上不同于服用这种蛋白质。基于 在这些观察中，申请人建议检验以下内容 假设：1)质粒载体的选择性甲基化可用于 通过延长转基因时间来提高转基因传递的效率 转基因表达。同样的甲基化也可以降低毒性。 原核生物来源的DNA在体外和体内的细胞内；2) α-1抗胰蛋白酶基因转移作为抗病毒药物的治疗潜力 抗炎治疗可以通过将基因传递到 选择性甲基化的质粒载体。此第一阶段建议书旨在 扩大和完善DNA甲基化在几个细胞中的观察 类型和活体内。另一个目标是，更具体地说，表明 A-1抗胰蛋白酶基因的传递和表达得到增强，并 质粒/阳离子脂质体转染剂的毒性降低 使用选择性甲基化的质粒。在第二阶段建议中， 该组织预计将扩大增强转基因的体内测试 携带a-1抗胰蛋白酶基因的载体检测对两种疾病的疗效 病毒性和非传染性肺炎性疾病作为下一步 确定临床用途。预计将推出两款最终产品：一 改良的体外转染剂和一种基于基因的抗病毒/抗病毒 发炎药。 建议的商业应用：不可用

DESCRIPTION (Adapted from investigator's abstract): The applicants have made two novel observations that have clear commercial potential. The first observation was the unexpected finding that methylated plasmid DNA extended the length of time a transgene is expressed in mammalian lung cells. This extension of expression is most likely due to retention of the transgene in the nucleus of transfected cells for a longer time than non-methylated plasmid DNA. The second observation is that expression of an a-1 antitrypsin transgene in lung cells has a therapeutic effect that is qualitatively different than administration of the protein. Based on these observations, the applicants propose to test the following hypotheses: 1) Selected methylation of plasmid vectors can be used to increase the efficiency of transgene delivery by extending the time of transgene expression. This same methylation can also reduce the toxicity of prokaryotic derived DNA in cells in vitro and in vivo; 2) The therapeutic potential of alpha-1 antitrypsin gene transfer as antiviral or anti-inflammatory therapy can be enhanced by delivering the gene in a selectively methylated plasmid vector. This phase I proposal is designed to extend and refine the DNA methylation observations in several cell types and in vivo. Another aim is to show, more specifically, that delivery and expression of the a-1 antitrypsin gene is enhanced and the toxicity of plasmid/cationic liposome transfection is decreased by employing a selectively methylated plasmid. In Phase II proposal the group anticipates extending the in vivo testing of the enhanced transgene vector carrying the a-1 antitrypsin gene to determine efficacy in both viral and non-infectious lung inflammatory disease as a next step toward determining clinical utility. Two eventual products are anticipated: an improved in vitro transfection reagent and a gene-based antiviral/anti- inflammatory drug. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE