Gene editing and elimination of latent herpes simplex virus in vivo
Gene editing and elimination of latent herpes simplex virus in vivo
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DOI：
10.1038/s41467-020-17936-5
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发表时间：
2020-08-18
影响因子：
16.6
通讯作者：
Jerome, Keith R.
中科院分区：
文献类型：
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作者：
Aubert, Martine;Strongin, Daniel E.;Jerome, Keith R.
我们使用腺相关病毒（AAV）递送的大范围核酸酶在一个成熟的小鼠模型中评估HSV的基因编辑，作为治疗潜伏性HSV感染的潜在治愈方法。在这里，我们证明了AAV递送的大范围核酸酶，而不是CRISPR/Cas9，介导了HSV的高效基因编辑，从上级颈神经节中消除了超过90%的潜伏病毒。单细胞RNA测序表明HSV和单个AAV血清型均非随机地分布在神经节中的神经元亚群中，这意味着向所有神经元亚群的改善的递送可导致HSV的甚至更完全的消除。如所预测的，使用三重AAV血清型组合递送大范围核酸酶导致神经节HSV负荷的最大降低。这些研究中观察到的HSV消除水平，如果转化为人类，可能会显著减少HSV再激活、脱落和病变。大范围核酸酶递送和活性的进一步优化很可能是可能的，并且可以提供治愈HSV感染的途径。
We evaluate gene editing of HSV in a well-established mouse model, using adeno-associated virus (AAV)-delivered meganucleases, as a potentially curative approach to treat latent HSV infection. Here we show that AAV-delivered meganucleases, but not CRISPR/Cas9, mediate highly efficient gene editing of HSV, eliminating over 90% of latent virus from superior cervical ganglia. Single-cell RNA sequencing demonstrates that both HSV and individual AAV serotypes are non-randomly distributed among neuronal subsets in ganglia, implying that improved delivery to all neuronal subsets may lead to even more complete elimination of HSV. As predicted, delivery of meganucleases using a triple AAV serotype combination results in the greatest decrease in ganglionic HSV loads. The levels of HSV elimination observed in these studies, if translated to humans, would likely significantly reduce HSV reactivation, shedding, and lesions. Further optimization of meganuclease delivery and activity is likely possible, and may offer a pathway to a cure for HSV infection.