CRISPR/Cas9 technology toward understanding HIV latency in primary cells

在接受高效联合抗逆转录病毒治疗的个体中潜伏HIV的存在 （cART）已经建立。如果没有额外的治疗，个人必须无限期地继续接受ART， 以避免出现严重的免疫缺陷和病毒传播。由于存在 治疗中断总是导致病毒反弹。因此，人们普遍认为， 根除病毒需要消除潜伏的宿主。 该提案的基本目标是调整我们的主要基于中医药的HIV-1潜伏期检测 在一些实施方案中，上文所述的用于通过CRISPR/Cas9技术测试潜伏期调节基因的方法是可行的。我们 因此，最初的靶点将是被广泛接受的候选基因，这些候选基因对以下两个方面都很重要： 延迟的建立/维持或从延迟中重新激活，以及一些以前未测试的 这些基因是通过不同实验室的基因筛选鉴定出来的。 控制病毒潜伏期的新基因/蛋白质的成功验证将导致识别 相关的途径和机制研究，将有可能产生重大的新举措， 开发新的治疗方法。

The presence of latent HIV in individuals treated with highly active combination antiretroviral therapy (cART) has been well established. Without additional therapies, individuals must remain on ART indefinitely in order to avoid development of severe immunodeficiency and spread of the virus. Because of the existence of this reservoir, treatment interruption invariably leads to viral rebound. Thus, it is generally accepted that eradication of the virus will require elimination of the latent reservoir. The fundamental goal of this proposal is adapt our primary TCM-based assay for HIV-1 latency described above to be amenable for testing of latency regulator genes via CRISPR/Cas9 technology. Our initial targets will therefore be candidate genes that are widely accepted to be important for either the establishment/maintenance of latency or for reactivation from latency, and a number of previously untested genes that were identified through genetic screens in various laboratories. Successful validation of novel genes/proteins that control viral latency will lead to identification of relevant pathways and mechanistic studies that will have the potential to spawn major new initiatives to develop novel therapeutics.