利用 CRISPR/Cas9 系统作为人类细胞内抵御 HIV-1 感染的防御机制。

Use of the CRISPR/Cas9 system as an intracellular defense against HIV-1 infection in human cells.

机构信息

Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.

Nomis Center for Immunobiology and Microbial Pathogenesis, Salk Institute for Biological Studies, La Jolla, California 92037, USA.

出版信息

Nat Commun. 2015 Mar 10;6:6413. doi: 10.1038/ncomms7413.

DOI:10.1038/ncomms7413

PMID:25752527

Abstract

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.

摘要

为了对抗有害病毒、细菌和古菌已经进化出一种独特的抗病毒防御系统，该系统由成簇规律间隔短回文重复序列（CRISPRs）和 CRISPR 相关基因（Cas）组成。CRISPR/Cas9 系统通过在特定位置产生 DNA 双链断裂（DSBs），从而发展出针对外来质粒和病毒的适应性免疫抵抗能力。在这里，我们将 CRISPR/Cas9 系统适应于人类细胞，以实现对细胞内外源 DNA 和病毒的防御。我们使用 HIV-1 感染作为模型，结果表明 CRISPR/Cas9 系统可以破坏潜伏整合的病毒基因组，并为人类细胞中新型病毒感染、表达和复制提供长期的适应性防御。我们还表明，稳定表达靶向 HIV 的 CRISPR/Cas9 的工程化人诱导多能干细胞可以有效地分化为 HIV 储存细胞类型，并保持对 HIV-1 挑战的抗性。这些结果揭示了 CRISPR/Cas9 系统作为一种新型抗病毒感染治疗策略的潜力。