Novel RNA Duplex Locks HIV-1 in a Latent State via Chromatin-mediated Transcriptional Silencing.
Novel RNA Duplex Locks HIV-1 in a Latent State via Chromatin-mediated Transcriptional Silencing.
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DOI：
10.1038/mtna.2015.31
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发表时间：
2015-10-27
期刊：
影响因子：
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通讯作者：
Suzuki K
中科院分区：
文献类型：
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作者：
Ahlenstiel C;Mendez C;Lim ST;Marks K;Turville S;Cooper DA;Kelleher AD;Suzuki K
哺乳动物基因的转录基因沉默 (TGS) 可以通过靶向启动子区域的短干扰 RNA (siRNA) 来诱导。我们之前报道过 siRNA (PromA) 对 HIV-1 具有有效的 TGS，其靶向病毒 5'LTR 内的串联 NF-κB 基序。在这项研究中，我们筛选了 siRNA 组合，旨在识别新的 5'LTR 靶标，以提供增强病毒沉默的多重潜力以及用于开发替代治疗策略的应用。系统检查发现了一个新的 siRNA 靶标 si143，并证实其可诱导 TGS 作为沉默机制。尽管诱导 TGS 后的能力有限，但 TGS 随病毒抑制而延长 >12 天。组蛋白脱乙酰酶抑制剂部分逆转表明与沉默相关的表观遗传变化，并通过染色质免疫沉淀分析证实，显示 H3K27me3 和 H3K9me3 的诱导、H3K9Ac 的减少以及 argonaute-1 的募集，这些都是异染色质和 TGS 的特征标记。总之，这些表观遗传变化模仿了与 HIV-1 潜伏期相关的变化。此外，当用 shPromA 和/或 sh143 转导时，在 J-Lat 9.2 细胞潜伏期模型中观察到对再激活的强大抵抗力。这些数据支持 si/shRNA 介导的 HIV-1 TGS 方法，并提供了追求功能性治愈的替代目标，从而使病毒库在抗逆转录病毒治疗停止后被锁定在潜伏期。
Transcriptional gene silencing (TGS) of mammalian genes can be induced by short interfering RNA (siRNA) targeting promoter regions. We previously reported potent TGS of HIV-1 by siRNA (PromA), which targets tandem NF-κB motifs within the viral 5′LTR. In this study, we screened a siRNA panel with the aim of identifying novel 5′LTR targets, to provide multiplexing potential with enhanced viral silencing and application toward developing alternate therapeutic strategies. Systematic examination identified a novel siRNA target, si143, confirmed to induce TGS as the silencing mechanism. TGS was prolonged with virus suppression >12 days, despite a limited ability to induce post- TGS. Epigenetic changes associated with silencing were suggested by partial reversal by histone deacetylase inhibitors and confirmed by chromatin immunoprecipitation analyses, which showed induction of H3K27me3 and H3K9me3, reduction in H3K9Ac, and recruitment of argonaute-1, all characteristic marks of heterochromatin and TGS. Together, these epigenetic changes mimic those associated with HIV-1 latency. Further, robust resistance to reactivation was observed in the J-Lat 9.2 cell latency model, when transduced with shPromA and/or sh143. These data support si/shRNA-mediated TGS approaches to HIV-1 and provide alternate targets to pursue a functional cure, whereby the viral reservoir is locked in latency following antiretroviral therapy cessation.
DOI：
10.1038/mtna.2012.8
发表时间：
2012-04-10
期刊：
Molecular therapy. Nucleic acids
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通讯作者：
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