Antiviral Targeting of Varicella Zoster Virus Replication and Neuronal Reactivation Using CRISPR/Cas9 Cleavage of the Duplicated Open Reading Frames 62/71

Betty W. Wu, Michael B. Yee, Ronald S. Goldstein, Paul R. Kinchington

Research output: Contribution to journalArticlepeer-review

2 Scopus citations


Varicella Zoster Virus (VZV) causes Herpes Zoster (HZ), a common debilitating and complicated disease affecting up to a third of unvaccinated populations. Novel antiviral treatments for VZV reactivation and HZ are still in need. Here, we evaluated the potential of targeting the replicating and reactivating VZV genome using Clustered Regularly Interspaced Short Palindromic Repeat-Cas9 nucleases (CRISPR/Cas9) delivered by adeno-associated virus (AAV) vectors. After AAV serotype and guide RNA (gRNA) optimization, we report that a single treatment with AAV2-expressing Staphylococcus aureus CRISPR/Cas9 (saCas9) with gRNA to the duplicated and essential VZV genes ORF62/71 (AAV2-62gRsaCas9) greatly reduced VZV progeny yield and cell-to-cell spread in representative epithelial cells and in lytically infected human embryonic stem cell (hESC)-derived neurons. In contrast, AAV2-62gRsaCas9 did not reduce the replication of a recombinant virus mutated in the ORF62 targeted sequence, establishing that antiviral effects were a consequence of VZV-genome targeting. Delivery to latently infected and reactivation-induced neuron cultures also greatly reduced infectious-virus production. These results demonstrate the potential of AAV-delivered genome editors to limit VZV productive replication in epithelial cells, infected human neurons, and upon reactivation. The approach could be developed into a strategy for the treatment of VZV disease and virus spread in HZ.

Original languageEnglish
Article number378
Issue number2
StatePublished - 12 Feb 2022

Bibliographical note

Publisher Copyright:
© 2022 by the authors. Licensee MDPI, Basel, Switzerland.


Funding: This work was supported by an Israel-US Binational Science Foundation award (#2017259; PRK and RSG), awards from the National Institutes of Health from NIAID (AI122640), and a P30 CORE grant from NEI (EY08098). BWW was also supported by a predoctoral award under T32 AI049820 (N.A. DeLuca, PI). Acknowledgments: The authors wish to acknowledge unrestricted support from the Eye & Ear Foundation of Pittsburgh and the Research to Prevent Blindness Inc., NY. The authors also wish to acknowledge advice and assistance from Leah Byrne, on the preparation of AAV; and Kira Lathrop, MAMS, for assisting with image processing.

FundersFunder number
National Institutes of Health
National Eye InstituteT32 AI049820, EY08098
National Institute of Allergy and Infectious DiseasesR01AI122640
Eye and Ear Foundation of Pittsburgh
Research to Prevent Blindness
United States-Israel Binational Science Foundation2017259


    • AAV
    • Antiviral therapies
    • CRISPR/Cas9
    • Genome cleavage
    • Latency
    • Reactivation
    • Varicella zoster virus


    Dive into the research topics of 'Antiviral Targeting of Varicella Zoster Virus Replication and Neuronal Reactivation Using CRISPR/Cas9 Cleavage of the Duplicated Open Reading Frames 62/71'. Together they form a unique fingerprint.

    Cite this