Inhibition of HIV-1 infection using a micro-RNA cluster

使用 micro-RNA 簇抑制 HIV-1 感染

• 批准号: 7010704
• 负责人: DANIEL BODEN
• 金额: $ 21.97万
• 依托单位: AARON DIAMOND AIDS RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7010704
• 关键词: HIV infections; RNA interference; antiAIDS agent; bioengineering /biomedical engineering; biotechnology; blood banks; cell line; chemokine receptor; gene expression; gene targeting; host organism interaction; human immunodeficiency virus 1; human tissue; lymphocyte; transfection /expression vector; virus genetics; virus infection mechanism

DESCRIPTION (provided by applicant): The concept of using RNA molecules as therapeutic agents has recently received increased attention with the identification of RNA interference (RNAi). RNAi has emerged as a powerful tool to silence gene expression in multiple organisms. Recent research has demonstrated the utility of RNAi in inhibiting the replication of viruses such as HIV-1. Sequence-specific degradation of viral or cellular RNA has been achieved using small, dsRNA gene silencing triggers, termed small interfering RNAs (siRNAs). To introduce RNAi as a therapeutic modality against HIV-1 it must durably control the rapid replication kinetics and high degree of variation of HIV-1. We have previously demonstrated the emergence and selection of viral quasispecies resistant to siRNA targeting the HIV-1 transactivator protein tat. To counteract the rapid emergence of resistance, we hypothesize that co-expression of multiple siRNAs targeting conserved viral RNA sequences as well as mRNA sequences of critical host genes (e.g. CCR5) may reduce the emergence of siRNA resistant virus. The goal of this research proposal is to achieve intracellular antiretroviral immunity by DNA vectors, which express multiple siRNAs against HIV-1 RNA and co-receptor CCR5 delivered by a microRNA cluster. The specific aims of this study are: 1. To engineer pre-micro-siRNAs against HIV-1 and CCR5. 2. To generate AAV-DNA vectors co-expressing multiple pre-micro-siRNAs targeting different viral and host genes. 3. To determine the antiviral activity and durability of HIV-1 inhibition by the expression of multiple siRNAs in stable cell lines and primary lymphocytes.

描述（由申请人提供）：使用RNA分子作为治疗剂的概念最近通过鉴定RNA干扰（RNAI）而受到了越来越多的关注。 RNAi已成为一种强大的工具，可以使多种生物体中的基因表达沉默。最近的研究表明，RNAi在抑制了HIV-1等病毒的复制方面的实用性。使用小的DSRNA基因沉默触发器，已称为小型干扰RNA（siRNA），已经实现了病毒或细胞RNA的序列特异性降解。为了将RNAi作为针对HIV-1的治疗方式，它必须持久地控制快速复制动力学和HIV-1的高度变化。我们以前已经证明了抗HIV-1反式激活蛋白TAT的抗siRNA的病毒准确性的出现和选择。为了抵消耐药的快速出现，我们假设靶向保守的病毒RNA序列的多个siRNA的共表达以及关键宿主基因的mRNA序列（例如CCR5）可能会降低抗siRNA耐药性病毒的出现。该研究建议的目的是通过DNA载体实现细胞内抗逆转录病毒的免疫力，该载体对MicroRNA群集传递了针对HIV-1 RNA和共受体CCR5的多个siRNA。 这项研究的具体目的是： 1。用于针对HIV-1和CCR5的微丝前体。 2。为了产生AAV-DNA载体，共表达了靶向不同病毒和宿主基因的多个微肌中。 3。通过稳定细胞系和原发性淋巴细胞中多个siRNA的表达来确定HIV-1抑制的抗病毒活性和耐用性。