Impact of Tat-binding Cellular LncRNAs on HIV replication

摘要 非编码RNA(NcRNA)代表了人类转录组的大部分。长的非编码RNA (LncRNA)调节无数的细胞、发育、免疫反应和疾病相关过程。 并正在成为诊断生物标记物。最近的报告证明了细胞的重要性 关于艾滋病毒复制和潜伏期的lncRNA。LncRNAs与HIV蛋白相互作用，但这些蛋白的确切作用 病毒复制、潜伏和重新激活过程中的相互作用在很大程度上是未知的。已有研究表明，lncRNA可以 快速有效地调节其蛋白质结合伙伴的细胞运输、定位和功能。 LncRNAs在病毒感染时受到调节，因此它们可以迅速与病毒蛋白相互作用并影响病毒。 细胞内的复制周期。然而，细胞内的lncRNA与病毒蛋白直接相互作用的可能性 增强或干扰它们的功能并影响病毒复制还没有系统的探索。这个 已知HIV-Tat蛋白与细胞RNA和至少一种lncRNA NRON相互作用。在我们的预赛中 研究发现，TAT与细胞内lncRNAs之间存在20种新的相互作用。其中一种TAT装订 转移相关肺腺癌转录本1(MALAT1)抑制HIV在细胞内复制 体外感染原代CD4T细胞。我们假设MALAT1影响HIV 通过其与TAT的相互作用而感染。在这个建议中，我们寻求开发一种规则间隔的簇状 短回文重复干扰(CRISPRi)功能丧失筛查并确定TOP5的效果 TAT结合的细胞内lncRNAs对HIV复制的影响(Aim1)及其分子机制 抑制(AIM2)。这项拟议的研究将首次全面剖析Tat结合细胞的影响。 HIV上的IncRNA。已发现的影响HIV复制的lncRNAs可用作潜在的治疗靶点。 发现，产生的数据将扩展关于细胞内lncRNAs在艾滋病毒感染中的作用的当前信息 LncRNAs与HIV蛋白之间功能相关的新相互作用及其分子靶点的确定 抗艾滋病毒治疗。

Summary Non-coding RNAs (ncRNA) represent the majority of the human transcriptome. The long non-coding RNAs (LncRNA) regulate a myriad of cellular, developmental, immune response and disease-associated processes and are emerging as diagnostic biomarkers. Recent reports have demonstrated the importance of cellular LncRNAs on HIV replication and latency. LncRNAs interact with HIV proteins but the precise roles of these interactions in viral replication, latency, and reactivation are largely unknown. LncRNAs have been shown to swiftly and efficiently regulate cellular trafficking, localization, and function of their protein binding partners. LncRNAs are modulated upon viral infection, thus they can rapidly interact with viral proteins and influence viral replication cycle in the cells. However, the possibility that the cellular LncRNAs interact with viral proteins directly to enhance or interfere with their function and affect viral replication has not been explored systematically. The HIV-Tat protein is known to interact with cellular RNAs and at least one LncRNA, NRON. In our preliminary investigation, we found 20 novel interactions between Tat and cellular LncRNAs. One of the Tat binding LncRNAs, metastasis associated lung adenocarcinoma transcript 1 (MALAT1) inhibited HIV replication in a cell line model as well as ex vivo infected primary CD4+ T cells. We hypothesize that MALAT1 influences HIV infection through its interaction with Tat. In this proposal, we seek to develop a Clustered Regularly Interspaced Short Palindromic Repeats interference (CRISPRi) loss-of-function screen and determine the effects of top 5 Tat-binding cellular LncRNAs on HIV replication (Aim1) and molecular mechanisms of MALAT1 mediated HIV inhibition (Aim2). The proposed study will be the first to comprehensively dissect the impact of Tat binding cellular LncRNAs on HIV. The LncRNAs found to affect HIV replication can be used as potential therapeutic targets. The data generated will expand the current information on the role of cellular LncRNAs in HIV infection, discover functionally relevant novel interactions between LncRNAs and HIV proteins and identify molecular targets for anti-HIV therapy.