Zinc finger nuclease: a new approach for excising HIV-1 proviral DNA from infected human T cells
Zinc finger nuclease: a new approach for excising HIV-1 proviral DNA from infected human T cells
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DOI：
10.1007/s11033-014-3456-3
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发表时间：
2014-06
影响因子：
2.8
通讯作者：
Xiying Qu;Pengfei Wang;Donglin Ding;Xiaohui Wang;Gongmin Zhang;Xin Zhou;Lin Liu;Xiaoli Zhu
中科院分区：
文献类型：
--
作者：
Xiying Qu;Pengfei Wang;Donglin Ding;Xiaohui Wang;Gongmin Zhang;Xin Zhou;Lin Liu;Xiaoli Zhu
获得性免疫缺陷综合征（AIDS）不能完全治愈的一个主要原因是人类免疫缺陷病毒1 （HIV-1）原病毒被整合到人类基因组中。尽管现有的治疗方法可以抑制HIV-1的复制，但它们无法根除它。一种分子疗法因其特异性靶向HIV-1感染细胞并有效去除HIV-1而受到欢迎，无论病毒基因是活跃的还是休眠的。现在，我们提出了一种新的方法，可以很好地从被感染的人T细胞基因组中删除HIV原病毒。首先，我们设计了锌指核酸酶（ZFNs），其靶向长末端重复序列（LTR） U3区域内的一个序列，该序列在整个进化枝中高度保守。然后，我们筛选出一对ZFN，命名为ZFN- u3。我们发现ZFN-U3能准确靶向并清除被感染的人细胞系后的全长HIV-1前病毒DNA，其切除频率约为30%，且无细胞毒性。这些结果证明ZFN-U3可以有效地从感染细胞的人类基因组中剔除整合的HIV-1。因此，这种在人类基因组中删除全长HIV-1的方法可以为未来治愈hiv感染者提供一种新的方法。
A major reason that Acquired Immune Deficiency Syndrome (AIDS) cannot be completely cured is the human immunodeficiency virus 1 (HIV-1) provirus integrated into the human genome. Though existing therapies can inhibit replication of HIV-1, they cannot eradicate it. A molecular therapy gains popularity due to its specifically targeting to HIV-1 infected cells and effectively removing the HIV-1, regardless of viral genes being active or dormant. Now, we propose a new method which can excellently delete the HIV provirus from the infected human T cell genome. First, we designed zinc-finger nucleases (ZFNs) that target a sequence within the long terminal repeat (LTR) U3 region that is highly conserved in whole clade. Then, we screened out one pair of ZFN and named it as ZFN-U3. We discovered that ZFN-U3 can exactly target and eliminate the full-length HIV-1 proviral DNA after the infected human cell lines treated with it, and the frequency of its excision was about 30 % without cytotoxicity. These results prove that ZFN-U3 can effciently excise integrated HIV-1 from the human genome in infected cells. This method to delete full length HIV-1 in human genome can therefore provide a novel approach to cure HIV-infected individuals in the future.