Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells
Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells
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DOI：
10.1038/ncomms7413
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发表时间：
2015-03-01
影响因子：
16.6
通讯作者：
Belmonte, Juan Carlos Izpisua
中科院分区：
文献类型：
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作者：
Liao, Hsin-Kai;Gu, Ying;Belmonte, Juan Carlos Izpisua
为了对抗敌对病毒，细菌和古细菌进化出了一种独特的抗病毒防御系统，该系统由成簇的规则间隔的短回文重复序列 (CRISPR) 以及 CRISPR 相关基因 (Cas) 组成。 CRISPR/Cas9 系统通过产生位点特异性 DNA 双链断裂 (DSB)，对外来质粒和病毒产生适应性免疫抵抗力。在这里，我们将 CRISPR/Cas9 系统应用于人类细胞，用于细胞内防御外来 DNA 和病毒。使用 HIV-1 感染作为模型，我们的结果表明 CRISPR/Cas9 系统破坏了潜在整合的病毒基因组，并为人类细胞中的新病毒感染、表达和复制提供了长期的适应性防御。我们证明，稳定表达 HIV 靶向 CRISPR/Cas9 的工程化人类诱导多能干细胞可以有效分化为 HIV 储存细胞类型，并保持其对 HIV-1 攻击的抵抗力。这些结果揭示了 CRISPR/Cas9 系统作为针对病毒感染的新治疗策略的潜力。
To combat hostile viruses, bacteria and archaea have evolved a unique antiviral defense system composed of clustered regularly interspaced short palindromic repeats (CRISPRs), together with CRISPR-associated genes (Cas). The CRISPR/Cas9 system develops an adaptive immune resistance to foreign plasmids and viruses by creating site-specific DNA double-stranded breaks (DSBs). Here we adapt the CRISPR/Cas9 system to human cells for intracellular defense against foreign DNA and viruses. Using HIV-1 infection as a model, our results demonstrate that the CRISPR/Cas9 system disrupts latently integrated viral genome and provides long-term adaptive defense against new viral infection, expression and replication in human cells. We show that engineered human-induced pluripotent stem cells stably expressing HIV-targeted CRISPR/Cas9 can be efficiently differentiated into HIV reservoir cell types and maintain their resistance to HIV-1 challenge. These results unveil the potential of the CRISPR/Cas9 system as a new therapeutic strategy against viral infections.