IMPROVED NON VIRAL GENE TRANSFER VECTOR

描述（改编自研究者摘要）：申请人有 提出了两个具有明显商业潜力的新观察结果。 的 第一个观察结果是意外发现甲基化质粒DNA 延长转基因在哺乳动物肺中表达的时间长度 细胞 这种表达的延伸很可能是由于保留了 转基因在转染细胞的细胞核中的时间比 非甲基化质粒DNA。 第二个观察是， 肺细胞中的α-1抗胰蛋白酶转基因具有治疗效果， 与蛋白质的给药有质的不同。 基于 根据这些观察，申请人提出测试以下内容 假设：1）质粒载体的选择的甲基化可用于 通过延长转基因递送的时间来提高转基因递送的效率， 转基因表达 同样的甲基化也可以降低毒性 在体外和体内的细胞中的原核衍生的DNA; 2） α-1抗胰蛋白酶基因转移作为抗病毒或 抗炎治疗可以通过以一种 选择性甲基化质粒载体。 第一阶段提案旨在 为了扩展和完善在几个细胞中的DNA甲基化观察， 类型和体内。 另一个目的是更具体地表明， 增强了α-1抗胰蛋白酶基因的递送和表达， 质粒/阳离子脂质体转染的毒性降低， 使用选择性甲基化的质粒。 在第二阶段提案中， 该小组预计将扩大增强型转基因的体内测试， 载体携带的α-1抗胰蛋白酶基因，以确定在两种 病毒性和非感染性肺炎性疾病作为下一步 确定临床效用。 两个最终的产品预计： 改进的体外转染试剂和基于基因的抗病毒/抗- 消炎药 拟议商业应用：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