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Article

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

by
Min Ying
1,2,3,4,†,
Huadong Wang
2,3,4,*,†,
Tongtan Liu
2,3,
Zengpeng Han
1,2,3,4,
Kunzhang Lin
2,3,
Qing Shi
2,3,5,
Ning Zheng
1,
Tao Ye
6,
Huinan Gong
2,3,7 and
Fuqiang Xu
1,2,3,4,8,*
1
State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Key Laboratory of Magnetic Resonance in Biological Systems, Wuhan Center for Magnetic Resonance, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan 430071, China
2
Key Laboratory of Quality Control Technology for Virus-Based Therapeutics, Guangdong Provincial Medical Products Administration, NMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal Products, The Brain Cognition and Brain Disease Institute, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
3
Shenzhen Key Laboratory of Viral Vectors for Biomedicine, Shenzhen-Hong Kong Institute of Brain Science, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China
4
University of Chinese Academy of Sciences, Beijing 100049, China
5
College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China
6
Chinese Academy of Sciences Key Laboratory of Brain Connectome and Manipulation, Shenzhen Key Laboratory of Translational Research for Brain Diseases, The Brain Cognition and Brain Disease Institute, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen-Hong Kong Institute of Brain Science, Shenzhen 518055, China
7
Jiangsu Key Laboratory of Brain Disease and Bioinformation, College of Life Sciences, Xuzhou Medical University, Xuzhou 221004, China
8
Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Pathogens 2023, 12(6), 814; https://doi.org/10.3390/pathogens12060814
Submission received: 16 April 2023 / Revised: 30 May 2023 / Accepted: 6 June 2023 / Published: 7 June 2023
(This article belongs to the Special Issue Herpesvirus Infections and Antiviral Drugs)
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Abstract

Herpes simplex virus type 1 (HSV-1) is a leading cause of encephalitis and infectious blindness. The commonly used clinical therapeutic drugs are nucleoside analogues such as acyclovir. However, current drugs for HSV cannot eliminate the latent virus or viral reactivation. Therefore, the development of new treatment strategies against latent HSV has become an urgent need. To comprehensively suppress the proliferation of HSV, we designed the CLEAR strategy (coordinated lifecycle elimination against viral replication). VP16, ICP27, ICP4, and gD—which are crucial genes that perform significant functions in different stages of the HSV infection lifecycle—were selected as targeting sites based on CRISPR-Cas9 editing system. In vitro and in vivo investigations revealed that genome editing by VP16, ICP27, ICP4 or gD single gene targeting could effectively inhibit HSV replication. Moreover, the combined administration method (termed “Cocktail”) showed superior effects compared to single gene editing, which resulted in the greatest decrease in viral proliferation. Lentivirus-delivered CRISPR-Cas9/gRNA editing could effectively block HSV replication. The CLEAR strategy may provide new insights into the potential treatment of refractory HSV-1-associated diseases, particularly when conventional approaches have encountered resistance.
Keywords: herpes simplex virus; CRISPR-Cas9; gRNA; lentivirus; VP16; ICP27; ICP4; gD herpes simplex virus; CRISPR-Cas9; gRNA; lentivirus; VP16; ICP27; ICP4; gD

Share and Cite

MDPI and ACS Style

Ying, M.; Wang, H.; Liu, T.; Han, Z.; Lin, K.; Shi, Q.; Zheng, N.; Ye, T.; Gong, H.; Xu, F. CLEAR Strategy Inhibited HSV Proliferation Using Viral Vectors Delivered CRISPR-Cas9. Pathogens 2023, 12, 814. https://doi.org/10.3390/pathogens12060814

AMA Style

Ying M, Wang H, Liu T, Han Z, Lin K, Shi Q, Zheng N, Ye T, Gong H, Xu F. CLEAR Strategy Inhibited HSV Proliferation Using Viral Vectors Delivered CRISPR-Cas9. Pathogens. 2023; 12(6):814. https://doi.org/10.3390/pathogens12060814

Chicago/Turabian Style

Ying, Min, Huadong Wang, Tongtan Liu, Zengpeng Han, Kunzhang Lin, Qing Shi, Ning Zheng, Tao Ye, Huinan Gong, and Fuqiang Xu. 2023. "CLEAR Strategy Inhibited HSV Proliferation Using Viral Vectors Delivered CRISPR-Cas9" Pathogens 12, no. 6: 814. https://doi.org/10.3390/pathogens12060814

APA Style

Ying, M., Wang, H., Liu, T., Han, Z., Lin, K., Shi, Q., Zheng, N., Ye, T., Gong, H., & Xu, F. (2023). CLEAR Strategy Inhibited HSV Proliferation Using Viral Vectors Delivered CRISPR-Cas9. Pathogens, 12(6), 814. https://doi.org/10.3390/pathogens12060814

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