EQUINE LENTIVRAL VECTORS FOR GENE DELIVERY

用于基因传递的马慢病毒载体

• 批准号: 6607091
• 负责人: JOHN C OLSEN
• 金额: $ 19.74万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-04-01 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6607091
• 关键词: athymic mouse; biotechnology; capsid; equine infectious anemia virus; gene expression; gene therapy; human tissue; laboratory mouse; laboratory rat; respiratory epithelium; technology /technique development; tissue /cell culture; transfection /expression vector; virus genetics; virus protein; virus replication; xenotransplantation

The overall goal is to maximize the potential of retrovirus-mediated gene transfer to airway epithelium and other in vivo targets. To do this, it is proposed to develop a new retroviral gene transfer system, based upon equine infectious anemia virus (EVIAV). Like other lentivirus members of the retrovirus family, EIAV is capable of gene transfer to non-dividing cells. This proposal is focused on the following aspects of EIAV gene transfer vector development. (i) defining the encapsidation requirements of EIAV RNA to generate safe and efficient lentiviral packaging cells; (ii) maximizing the safety and expression of EIAV RNA to generate safe and efficient lentiviral packaging cells; (ii) maximizing the safety and expression of EIAV vectors in vector producing cells; and (iii) investigating the entry and persistence of expression in human models of well differentiated airway epithelia. The focus on EIAV RNA encapsidation includes identifying important cis- acting sequence elements important for assembling vector into virus particles. Sequences important for encapsidation will be removed from expression cassettes. The focus on expression of EIAV vectors in producer cells includes determining the role of the viral regulatory/accessory genes on producing vectors in heterologous cells. In addition the effect of modifying the vector to prevent synthesis of viral related proteins and the effect of disabling the viral transcription regulatory elements will be determined. The focus on entry includes experiments designed to expand the host range of lentiviral vectors to include the apical membrane of polarized epithelia. In addition the persistence of EIAV vector of EIAV vector expression be studied in well differentiated. Our overall hypothesis is that a safe retroviral system of efficiently delivering genes of quiescent cells will have broad application for in vivo and ex vivo gene transfer to treat diseases of the lungs and other organs.

总体目标是最大限度地发挥逆转录病毒介导的潜力 基因转移至气道上皮和其他体内靶标。要做的事 为此，建议开发一种新的逆转录病毒基因转移系统， 基于马传染性贫血病毒（EVIAV）。与其他慢病毒一样 EIAV 是逆转录病毒家族的成员，能够将基因转移到 不分裂的细胞。该提案主要关注以下几个方面 EIAV基因转移载体开发。 (i) 定义衣壳 EIAV RNA 生成安全高效慢病毒的要求 包装细胞； (ii) 最大化 EIAV RNA 的安全性和表达 产生安全高效的慢病毒包装细胞； (二) 最大限度地提高 EIAV 载体在载体生产中的安全性和表达 细胞； (iii) 调查表达的进入和持续性 在分化良好的气道上皮的人类模型中。 EIAV RNA 衣壳化的重点包括识别重要的顺式 对于将载体组装成病毒很重要的作用序列元件 颗粒。对于封装重要的序列将从中删除 表达盒。 对 EIAV 载体在生产细胞中表达的关注包括 确定病毒调节/辅助基因的作用 在异源细胞中产生载体。另外的效果是 修饰载体以防止病毒相关蛋白的合成， 禁用病毒转录调控元件的效果将 被确定。 进入的重点包括旨在扩展宿主的实验 一系列慢病毒载体，包括极化的顶膜 上皮细胞。另外 EIAV 载体的持久性 EIAV 载体 表达进行了良好分化的研究。 我们的总体假设是，一个安全、高效的逆转录病毒系统 传递静止细胞的基因将在以下方面具有广泛的应用 体内和离体基因转移治疗肺部疾病和其他疾病 器官。