利用 CRISPR/Cas13d 核酸酶系统高效抑制 HIV。
Efficient Inhibition of HIV Using CRISPR/Cas13d Nuclease System.
机构信息
Host-Pathogen Interaction Program, Texas Biomedical Research Institute, San Antonio, TX 78227, USA.
Disease Intervention and Prevention Program, Texas Biomedical Research Institute, San Antonio, TX 78227, USA.
出版信息
Viruses. 2021 Sep 16;13(9):1850. doi: 10.3390/v13091850.
Recently discovered Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas13 proteins are programmable RNA-guided ribonucleases that target single-stranded RNA (ssRNA). CRISPR/Cas13-mediated RNA targeting has emerged as a powerful tool for detecting and eliminating RNA viruses. Here, we demonstrate the effectiveness of CRISPR/Cas13d to inhibit HIV-1 replication. We designed guide RNAs (gRNAs) targeting highly conserved regions of HIV-1. RfxCas13d (CasRx) in combination with HIV-specific gRNAs efficiently inhibited HIV-1 replication in cell line models. Furthermore, simultaneous targeting of four distinct, non-overlapping sites in the HIV-1 transcript resulted in robust inhibition of HIV-1 replication. We also show the effective HIV-1 inhibition in primary CD4 T-cells and suppression of HIV-1 reactivated from latently infected cells using the CRISPR/Cas13d system. Our study demonstrates the utility of the CRISPR/Cas13d nuclease system to target acute and latent HIV infection and provides an alternative treatment modality against HIV.
最近发现的成簇规律间隔短回文重复序列（CRISPR）/Cas13 蛋白是可编程的 RNA 指导的核糖核酸酶，可靶向单链 RNA（ssRNA）。CRISPR/Cas13 介导的 RNA 靶向已成为检测和消除 RNA 病毒的强大工具。在这里，我们证明了 CRISPR/Cas13d 抑制 HIV-1 复制的有效性。我们设计了针对 HIV-1 高度保守区域的向导 RNA（gRNA）。RfxCas13d（CasRx）与 HIV 特异性 gRNA 结合，可有效抑制细胞系模型中的 HIV-1 复制。此外，同时靶向 HIV-1 转录本中的四个不同的、不重叠的位点，可导致 HIV-1 复制的强大抑制。我们还表明，使用 CRISPR/Cas13d 系统可有效抑制原代 CD4 T 细胞中的 HIV-1 感染，并抑制潜伏感染细胞中 HIV-1 的重新激活。我们的研究证明了 CRISPR/Cas13d 核酸酶系统靶向急性和潜伏性 HIV 感染的实用性，并为 HIV 提供了一种替代治疗模式。
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