Identifying host factors essential for HIV latency maintenance

 描述(申请人提供)：高效抗逆转录病毒疗法(HAART)为患者提供长期抑制艾滋病毒复制，但它不能治愈患者，因为长期潜伏的艾滋病毒蓄水池-治愈艾滋病毒感染的主要障碍之一。潜伏艾滋病毒的重新激活一直在深入调查中；然而，由于对潜伏时间如何维持的了解有限，尚未制定具体重新激活潜伏艾滋病毒的战略。我们建议进行全基因组基因敲除筛选，以确定对HIV潜伏期维持至关重要的宿主因素，并全面了解其机制。直到最近出现了四篇描述CRISPR-Cas9介导的全基因组基因敲除筛选的论文，才有了在培养细胞中进行全基因组功能丧失筛选的方法。我们独立开发了一种类似的方法来进行全基因组筛选，以确定西尼罗河病毒诱导的细胞死亡所必需的宿主因子，结果表明，我们的方法可以识别具有高度特异性的靶基因。因此，我们有信心能够系统地识别宿主因素，以全面了解宿主因素如何促进HIV潜伏期维持的机制。我们将在不同的J-Lat克隆中测试已识别的基因，以确定可能特异性和普遍性地重新激活潜伏HIV的宿主因素，这可能会导致一种新的治疗方法来消除潜伏的HIV储备库。

 DESCRIPTION (provided by applicant): Highly active antiretroviral therapy (HAART) offers patients long-term suppression of HIV replication, but it does not cure the patients because of the long lasting latent HIV reservoir- one of the major obstacles in curing HIV infection. Reactivation of latent HIV has been under intensive investigation; however, no strategy has been developed to specifically reactivate latent HIV due to limited knowledge of how the latency is maintained. We propose to perform genome- wide knockout screening to identify the host factors that are essential for HIV latency maintenance and gain comprehensive insight into the mechanism. There has been no method for performing genome-wide loss-of-function screening in cultured cells until the appearance of four recent papers that described a CRISPR-Cas9-mediated genome-wide knockout screen. We have independently developed a similar approach to perform genome-wide screening to identify the host factors that are indispensable for WNV-induced cell death, and the results show that our method can identify target genes with high specificity. Thus, we are confident that we can identify the host factors systematically to gain comprehensive insight into the mechanism of how host factors facilitate HIV latency maintenance. We will test the identified genes in different J-Lat clones to identify the host factors that might reactivate latent HIV specifically and universally, which might lead to a new therapy to eradicate latent HIV reservoir.