Inhibition of human immunodeficiency virus type 1 replication with artificial transcription factors targeting the highly conserved primer-binding site
Inhibition of human immunodeficiency virus type 1 replication with artificial transcription factors targeting the highly conserved primer-binding site
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DOI：
10.1128/jvi.80.6.2873-2883.2006
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发表时间：
2006-03-01
影响因子：
5.4
通讯作者：
Barbas, CF
中科院分区：
文献类型：
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作者：
Eberhardy, SR;Goncalves, J;Barbas, CF
人类免疫缺陷病毒1型（HIV - 1）引物结合位点（PBS）是HIV基因组中一个高度保守的区域，是开发新型抗HIV疗法的一个有吸引力的靶点。在这项研究中，我们设计了四个人工锌指转录因子，使其结合在PBS处或其附近，并抑制HIV - 1长末端重复序列（LTR）的转录。染色质免疫沉淀分析表明，这些蛋白质在体内与LTR结合。在瞬时报告基因检测中，四种蛋白质中有三种抑制了由HIV - 1 LTR驱动的报告基因的转录。然而，只有一种被命名为KRAB - PBS2的蛋白质在转导到原代淋巴细胞中时能够阻止病毒产生。与未转导的细胞相比，我们观察到在数周的过程中病毒复制被抑制了>90%，并且没有观察到明显的细胞毒性。HIV - 1长期暴露于KRAB - PBS2会诱导HIV - 1 PBS发生突变，从而降低抑制因子的有效性，但这些突变也导致病毒复制速率下降。这些结果表明，KRAB - PBS2有潜力用于艾滋病患者的抗病毒治疗，并可能补充其他基于基因的策略。
The human immunodeficiency virus type 1 (HIV-1) primer-binding site (PBS) is a highly conserved region in the HIV genome and represents an attractive target for the development of new anti-HIV therapies. In this study, we designed four artificial zinc finger transcription factors to bind at or adjacent to the PBS and repress transcription from the HIV-1 long terminal repeat (LTR). These proteins bound to the LTR in vivo, as demonstrated by the chromatin immunoprecipitation assay. In transient reporter assays, three of the four proteins repressed transcription of a reporter driven by the HIV-1 LTR. Only one of these proteins, however, designated KRAB-PBS2, was able to prevent virus production when transduced into primary lymphocytes. We observed > 90% inhibition of viral replication over the course of several weeks compared to untransduced cells, and no significant cytotoxicity was observed. Long-term exposure of HIV-1 to KRAB-PBS2 induced mutations in the HIV-1 PBS that reduced the effectiveness of the repressor, but these mutations also resulted in decreased rates of viral replication. These results show that KRAB-PBS2 has the potential to be used in antiviral therapy for AIDS patients and might complement other gene-based strategies.