ANTISENSE RNA INHIBITION OF HIV VIA RETROVIRAL VECTORS

反义 RNA 通过逆转录病毒载体抑制 HIV

• 批准号: 3143324
• 负责人: Eli Gilboa
• 金额: $ 23.47万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1992-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3143324
• 关键词: AIDS therapy; DNA directed RNA polymerase; RNA biosynthesis; Retroviridae; antisense nucleic acid; antiviral agents; bacteriophage T7; biotechnology; gel electrophoresis; gene expression; genetic manipulation; genetic promoter element; human immunodeficiency virus; lymphocyte; murine leukemia virus; nucleic acid hybridization; oncogenes; tissue /cell culture; transfer RNA; transposon /insertion element; virus replication

The objective of the proposed studies is to investigate the use of antisense RNA inhibition as a method to render cells resistant to infection and replication of human immunodeficiency virus (HIV). The experimental strategy involves stable gene transfer of DNA constructs, which serve as templates for the constitutive expression of antisense RNA in the transduced cells. This approach, also termed "intracellular immunization", contrasts with the use antisense oligonucleotides. The proposed studies will focus on the design of HIV specific antisense DNA templates which will be introduced into cultured human lymphoid cell lines via retroviral vectors, and tested for their ability to inhibit the replication of HIV. The major objective of the proposed studies is to design vector systems leading to efficient expression of antisense RNA in the target cells by exploring (a) the use of three types of promoters for antisense RNA expression: mammalian RNA polymerase II, human t-RNA (pol III), and a bacteriophage T7 promoter, (b) the use of two retroviral vector systems: the previously characterized N2 vector, and double copy DC vectors, a novel type of vector which was designed to improve the expression of transduced genes. Antisense RNA inhibition mediated via stable gene transfer will be used to study the mechanism of molecular and genetic regulation of HIV. A long term objective of these studies is to develop a treatment for AIDS in which bone marrow cells derived from a patient are rendered resistant to HIV replication. Development of improved antisense RNA technologies will be particularly useful to study the mechanism of action of recessive oncogenes.

拟议研究的目的是调查 反义RNA抑制作为使细胞抵抗 人类免疫缺陷病毒（HIV）感染和复制。 实验策略包括DNA的稳定基因转移 构建体，其充当组成型结构的模板。 反义RNA在转导细胞中的表达。 这 方法，也称为“细胞内免疫”，与 使用反义寡核苷酸。 拟议的研究将侧重于设计艾滋病毒特异性 反义DNA模板将被引入到培养的 人淋巴细胞系，并测试了 他们抑制艾滋病毒复制的能力。 主要 本研究的目的是设计向量系统 导致反义RNA在靶细胞中的有效表达 （a）使用三种类型的启动子， 反义RNA表达：哺乳动物RNA聚合酶II，人t-RNA (pol III）和噬菌体T7启动子，（B）使用两个 逆转录病毒载体系统：先前表征的N2载体， 和双拷贝DC载体，一种新型载体， 旨在提高转导基因的表达。 反义 通过稳定的基因转移介导的RNA抑制将用于 研究HIV的分子和遗传调控机制。 这些研究的长期目标是开发一种治疗方法， 对于艾滋病，其中来自患者的骨髓细胞被 使其对艾滋病毒复制具有抵抗力。 开发改进的反义RNA技术将是 对研究隐性遗传的作用机制特别有用。 致癌基因