Cell Type-Specific Gene Delivery in Mammalian Retina

哺乳动物视网膜中细胞类型特异性基因传递

• 批准号: 6955008
• 负责人: VIJAY P SARTHY
• 金额: $ 14.85万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6955008
• 关键词: Muller' s cell; Retroviridae; avian leukosis virus; biotechnology; gene delivery system; gene expression; gene therapy; genetically modified animals; glial fibrillary acidic protein; laboratory mouse; retina; transfection /expression vector; virus receptors

DESCRIPTION (provided by applicant): The development of vectors for cell-specific delivery is of considerable interest for many cell biological studies. The present proposal describes a cell targeting strategy based on the use of the receptor for subgroup-A avian leukosis virus, TVA, to target retroviral vectors carrying genes of interest to a specific cell type in the mouse retina. The proposed studies are based on the idea that expression of the avian retroviral receptor, TVA, under a Muller cell-specific promoter renders Muller cells selectively susceptible to infection with the avian leukosis virus, and any gene carried by the infecting virus is expressed only in Muller cells. Therefore, the TVA-based retroviral system provides a novel opportunity to selectively and efficiently target the delivery and expression of any gene of interest in the Muller cell. Specifically, the proposed experiments will investigate whether GFAP promoter can be used to selectively expressing TVA in Muller cells. Subsequent studies wiill examine whether intraocular injection of the retrovirus, HIV GFP(ALSV-A), in GFAP-tv-a transgenic mice, results in Green Fluorescent Protein (GFP) expression specifically in Muller cells. Although the focus of the present proposal is to target genes to Muller cells, the TVA-based system has a much broader scope in retinal cell and molecular biological research. With a judicious choice of cell typespecific promoter, it should be possible to selectively and efficiently target the delivery and expression of any gene of interest to a specific retinal cell type. In summary, the TVA-system offers a novel, exciting approach for delivering genes of interest to specific retinal cells, and can be expected to have a major impact on future studies in retinal cell biology.

描述（由申请人提供）：细胞特异性递送载体的开发对于许多细胞生物学研究具有相当大的意义。本发明描述了一种细胞靶向策略，其基于使用A亚群禽白血病病毒TVA的受体来将携带感兴趣基因的逆转录病毒载体靶向小鼠视网膜中的特定细胞类型。拟议的研究基于这样的想法：在穆勒细胞特异性启动子下表达鸟类逆转录病毒受体TVA，使穆勒细胞选择性地对鸟类白血病病毒感染易感，并且感染病毒携带的任何基因仅在穆勒细胞中表达。因此，基于TVA的逆转录病毒系统提供了一个新的机会，以选择性和有效地靶向穆勒细胞中任何感兴趣的基因的递送和表达。具体地，所提出的实验将研究GFAP启动子是否可以用于在Muller细胞中选择性地表达TVA。随后的研究将检查在GFAP-tv-a转基因小鼠中眼内注射逆转录病毒HIV GFP（ALSV-A）是否导致绿色荧光蛋白（GFP）在Muller细胞中特异性表达。虽然本提案的重点是将基因靶向Muller细胞，但基于TVA的系统在视网膜细胞和分子生物学研究中具有更广泛的范围。通过明智地选择细胞类型特异性启动子，应该可以选择性地和有效地将任何感兴趣的基因的递送和表达靶向特定的视网膜细胞类型。总之，TVA系统提供了一种新的，令人兴奋的方法，用于将感兴趣的基因传递到特定的视网膜细胞，并有望对视网膜细胞生物学的未来研究产生重大影响。