Applications of CRISPR-Cas9 in the Treatment of Herpes Simplex Keratitis
DOI:
https://doi.org/10.54097/y1zmvr07Keywords:
HSK, HSV-1, HELP, CRISPR-Cas9, antiviral therapy.Abstract
Herpes simplex keratitis (HSK) is a leading cause of infectious corneal blindness. Herpes simplex virus type 1 (HSV-1) establishes lifelong latency in trigeminal ganglion neurons and reactivates in the cornea. While antivirals such as acyclovir (ACV) suppress viral replication, they do not eradicate the infection, and drug resistance is increasing. Genome editing offers a direct strategy to disrupt essential viral functions. CRISPR-Cas9, guided by single guide RNA (sgRNA), has been shown to reduce HSV-1 replication in cell cultures, organoids, and animal models, with multi-target designs limiting viral escape. Recent delivery advancements include HSV-1-erasing lentiviral particles (HELP), which package spCas9 mRNA with guides targeting UL8 and UL29, enabling transient editing in the cornea and retrograde transport to the trigeminal ganglion. Complementary approaches utilize adeno-associated virus serotype 2 (AAV2) to deliver SaCas9 targeting ICP0 and ICP27 in human induced pluripotent stem cell-derived cerebral organoids. Sensor-effector platforms such as autonomous virus-inducible immune-like cells (ALICE) integrate CRISPR editing with neutralizing antibodies, demonstrating antiviral activity comparable to high-dose ACV in certain contexts. Key challenges include off-target effects, access to latent chromatin, immunogenicity, and scalable ocular delivery. This review aims to synthesize current evidence, compare delivery platforms including HELP, AAV2, and ALICE, and identify priorities for clinical translation.
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