Cell Type-Specific Gene Delivery in Mammalian Retina

描述（由申请人提供）：细胞特异性传递载体的发展对许多细胞生物学研究具有相当大的兴趣。本提案描述了一种基于使用a亚群禽白血病病毒（TVA）受体的细胞靶向策略，以靶向携带小鼠视网膜中特定细胞类型感兴趣的基因的逆转录病毒载体。拟议的研究基于这样一种观点，即在Muller细胞特异性启动子下禽类逆转录病毒受体TVA的表达使Muller细胞选择性地易受禽类白血病病毒的感染，并且感染病毒所携带的任何基因仅在Muller细胞中表达。因此，基于tva的逆转录病毒系统提供了一种新的机会，可以选择性和有效地靶向Muller细胞中任何感兴趣的基因的传递和表达。具体来说，本实验将研究GFAP启动子是否可以在Muller细胞中选择性表达TVA。后续的研究将检验在GFAP-tv-a转基因小鼠眼内注射逆转录病毒HIV GFP（ALSV-A）是否会导致Muller细胞特异性表达绿色荧光蛋白（GFP）。虽然目前的研究重点是将基因靶向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应用范围。通过对细胞类型特异性启动子的明智选择，应该可以选择性和有效地靶向任何对特定视网膜细胞类型感兴趣的基因的传递和表达。总之，tva系统提供了一种新颖的、令人兴奋的方法，可以将感兴趣的基因传递到特定的视网膜细胞，并且可以预期对视网膜细胞生物学的未来研究产生重大影响。

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.