Herpes simplex virus type 1 (HSV-1) is a leading cause of infectious blindness. Current treatments for HSV-1 do not eliminate the virus from the site of infection or latent reservoirs in the trigeminal ganglia. Here, we target HSV-1 genomes directly using mRNA-carrying lentiviral particles that simultaneously deliver SpCas9 mRNA and viral-gene-targeting guide RNAs (designated HSV-1-erasing lentiviral particles, termed HELP). We show that HELP efficiently blocks HSV-1 replication and the occurrence of herpetic stromal keratitis (HSK) in three different infection models. HELP was capable of eliminating the viral reservoir via retrograde transport from corneas to trigeminal ganglia. Additionally, HELP inhibited viral replication in human-derived corneas without causing off-target effects, as determined by whole-genome sequencing. These results support the potential clinical utility of HELP for treating refractory HSK.

单纯疱疹病毒 1 型 (HSV-1) 是传染性失明的主要原因。目前对 HSV-1 的治疗不能从感染部位或三叉神经节中的潜伏宿主中消除病毒。在这里，我们使用携带 mRNA 的慢病毒颗粒直接靶向 HSV-1 基因组，这些颗粒同时递送 SpCas9 mRNA 和病毒基因靶向引导 RNA（指定 HSV-1 擦除慢病毒颗粒，称为 HELP）。我们展示了 HELP 在三种不同的感染模型中有效地阻止了 HSV-1 复制和疱疹性基质角膜炎 (HSK) 的发生。HELP 能够通过从角膜到三叉神经节的逆行运输来消除病毒库。此外，正如全基因组测序所确定的，HELP 抑制了人源角膜中的病毒复制，而不会引起脱靶效应。