CLEAR Strategy Inhibited HSV Proliferation Using Viral Vectors Delivered CRISPR-Cas9

Area of Science:

• Virology
• Gene Editing
• Infectious Diseases

Background:

• Herpes simplex virus type 1 (HSV-1) causes encephalitis and blindness.
• Current treatments like acyclovir fail to eliminate latent HSV or prevent reactivation.
• New strategies are urgently needed to combat latent HSV and drug resistance.

Purpose of the Study:

• To develop a novel gene-editing strategy for comprehensive HSV-1 suppression.
• To evaluate the efficacy of targeting crucial HSV-1 genes using CRISPR-Cas9.
• To assess the combined effect of targeting multiple HSV-1 genes.

Main Methods:

• Designed the CLEAR strategy targeting essential HSV-1 genes (VP16, ICP27, ICP4, gD) using CRISPR-Cas9.
• Conducted in vitro and in vivo experiments to assess single and combined gene editing efficacy.
• Utilized lentivirus delivery for CRISPR-Cas9/gRNA editing.

Main Results:

• Single gene targeting effectively inhibited HSV replication.
• The combined administration method ('Cocktail') showed superior antiviral effects compared to single gene editing.
• Lentivirus-delivered CRISPR-Cas9/gRNA effectively blocked HSV replication.

Conclusions:

• The CLEAR strategy provides a promising approach for treating HSV-1 infections.
• Combined gene editing offers enhanced efficacy against viral proliferation.
• This strategy holds potential for treating refractory HSV-1 diseases resistant to conventional therapies.