Single AAV-Mediated CRISPR-SaCas9 Inhibits HSV-1 Replication by Editing ICP4 in Trigeminal Ganglion Neurons
Single AAV-Mediated CRISPR-SaCas9 Inhibits HSV-1 Replication by Editing ICP4 in Trigeminal Ganglion Neurons
复制标题
单个 AAV 介导的 CRISPR-SaCas9 通过编辑三叉神经节神经元中的 ICP4 抑制 HSV-1 复制。
DOI：
10.1016/j.omtm.2020.05.011
复制
发表时间：
2020-09-11
影响因子：
4.7
通讯作者：
Zhou Songyang
中科院分区：
文献类型：
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作者：
Chen, Yuxi;Zhi, Shengyao;Zhou Songyang
单纯疱疹性角膜炎 (HSK) 是发达国家角膜失明的最常见原因，由角膜原发性或复发性单纯疱疹病毒 1 (HSV-1) 感染引起。 HSV-1的潜伏感染，尤其是三叉神经节（TG）中的潜伏感染，会导致HSV-1感染的复发。由于抗病毒治疗对潜伏的HSV-1无效，为了测试SpCas9（化脓性链球菌Cas9）或SaCas9（金黄色葡萄球菌Cas9）抑制HSV-1的可能性，选择了HSV-1复制和重新激活所需的两个重要基因ICPO和ICP4作为靶标。在Vero细胞中，靶向ICPO或ICP4的SpCas9和SaCas9可以有效抑制HSV-1的增殖，而不影响细胞活力。通过双基因组测序和深度测序验证，在人类基因组中没有观察到明显的向导RNA（gRNA）依赖性脱靶。腺相关病毒 1 (AAV1) 介导的 SaCas9 递送通过靶向小鼠原代 TG 神经元细胞中的 ICP4 抑制 HSV-1 复制。 SpCas9和SaCas9能够抑制Vero细胞和小鼠TG神经元培养物中的HSV-1感染，具有高效率和良好的生物安全性。 AAV1 介导的 SaCas9 递送在治疗 HSK 和抑制 TG 神经元中的 HSV-1 方面显示出巨大的潜力。可能需要进一步的研究来测试对潜伏感染的抑制，这可能会导致治疗病毒性疾病的新方法的开发。
Herpes simplex keratitis (HSK) is the most common cause of corneal blindness in developed nations, caused by primary or recurrent herpes simplex virus 1 (HSV-1) infection of the cornea. Latent infection of HSV-1, especially in the trigeminal ganglion (TG), causes recurrence of HSV-1 infection. As antiviral treatment is not effective on latent HSV-1, to test the possibility of inhibiting HSV-1 by SpCas9 (Streptococcus pyogenes Cas9) or SaCas9 (Staphylococcus aureus Cas9), ICPO and ICP4, two important genes required for HSV-1 replication and reactivation, were chosen as targets. In Vero cells, SpCas9 and SaCas9 targeting ICPO or ICP4 can effectively inhibit the proliferation of HSV-1 without affecting cell viability. No significant guide RNA (gRNA)-dependent off-targets were observed in the human genome by digenome sequencing and deep sequencing verification. Adeno-associated virus 1 (AAV1)-mediated delivery of SaCas9 inhibits HSV-1 replication by targeting ICP4 in mouse primary TG neuronal cells. SpCas9 and SaCas9 are able to inhibit HSV-1 infection in Vero cells and mouse TG neuronal cultures with high efficiency and good biosafety. AAV1-mediated delivery of SaCas9 shows great potential in treating HSK and inhibiting HSV-1 in TG neurons. Further investigations may be needed to test the inhibition of latent infections, which may result in the development of novel methods for treating viral diseases.