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.

中文翻译：

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使用 CRISPR/Cas13d 核酸酶系统有效抑制 HIV

最近发现的成簇规则间隔短回文重复序列 (CRISPR)/Cas13 蛋白是靶向单链 RNA (ssRNA) 的可编程 RNA 引导的核糖核酸酶。CRISPR/Cas13 介导的 RNA 靶向已成为检测和消除 RNA 病毒的有力工具。在这里，我们展示了 CRISPR/Cas13d 抑制 HIV-1 复制的有效性。我们设计了针对 HIV-1 高度保守区域的引导 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 转录物中四个不同的、不重叠的位点会导致对 HIV-1 复制的强烈抑制。我们还在原发性 CD4 ^{+中显示了有效的 HIV-1 抑制}使用 CRISPR/Cas13d 系统从潜伏感染的细胞中重新激活 T 细胞和抑制 HIV-1。我们的研究证明了 CRISPR/Cas13d 核酸酶系统在靶向急性和潜伏 HIV 感染方面的效用，并提供了一种针对 HIV 的替代治疗方式。