Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems

Abstract

Almost all people become infected with herpes viruses, including herpes simplex virus type 1 (HSV-1), during their lifetime. Typically, these viruses persist in a latent form that is resistant to all available antiviral medications. Under certain conditions, such as immunosuppression, the latent forms reactivate and cause disease. Moreover, strains of herpesviruses that are drug-resistant have rapidly emerged. Therefore, it is important to develop alternative methods capable of eradicating herpesvirus infections. One promising direction is the development of CRISPR/Cas systems for the therapy of herpesvirus infections. We aimed to design a CRISPR/Cas system for relatively effective long-term and safe control of HSV-1 infection. Here, we show that plasmids encoding the CRISPR/Cas9 system from Streptococcus pyogenes with a single sgRNA targeting the UL30 gene can completely suppress HSV-1 infection of the Vero cell line within 6 days and provide substantial protection within 9 days. For the first time, we show that CRISPR/CasX from Deltaproteobacteria with a single guide RNA against UL30 almost completely suppresses HSV-1 infection of the Vero cell line for 3 days and provides substantial protection for 6 days. We also found that the Cas9 protein without sgRNAs attenuates HSV-1 infection. Our results show that the developed CRISPR/Cas systems are promising therapeutic approaches to control HSV-1 infections.

Not Only Editing: A Cas-Cade of CRISPR/Cas-Based Tools for Functional Genomics in Plants and Animals

“…Numerous studies have already demonstrated the downregulation of viral infection in animal cell cultures. These studies have employed various types of Cas proteins depending on whether the targeted virus is based on DNA or RNA [ 91 , 92 ]. For example, CRISPR/Cas9 and Cas12a were shown to downregulate the infection of Green Monkey kidney cell culture by Herpes Simplex Virus 1 (HSV-1) [ 91 ].…”

“…These studies have employed various types of Cas proteins depending on whether the targeted virus is based on DNA or RNA [ 91 , 92 ]. For example, CRISPR/Cas9 and Cas12a were shown to downregulate the infection of Green Monkey kidney cell culture by Herpes Simplex Virus 1 (HSV-1) [ 91 ]. In this study, the authors targeted the HSV-1 genome with a Cas9 or Cas12a protein and a gRNA specific to a viral DNA polymerase and were able to see a clear diminution of the infection of the cell culture as well as an immunity that lasted for several days.…”

Viral delivery of CRISPR/Cas9 genome editing for rapid crop improvement: A promising approach to enhance crop resilience against biotic and abiotic stresses

“…Single-stranded RNA virus CRISPR/Cas9 system (K. Li et al, 2020) [67] Herpes Simplex Virus (HSV) Double-stranded DNA virus CRISPR/Cas9 system (Karpov et al, 2022) [59] Adeno-associated virus (AAV) Single-stranded DNA virus CRISPR/Cas9 with chemically modified sgRNA (Kim et al, 2021) [61]…”

L-lysine as a Possible Supplement for Treatment of Herpetic Epithelial Keratitis: A Case Report and Literature Review

“…15 Other researchers reinforce the importance of developing alternative methods for controlling these viruses and even capable of eradicating latent infections. 9,16, 17 Although current antivirals are still viable treatments for active herpetic diseases such as keratitis, controlling recurrence and the virus in the latent phase remains a challenge due to a virus in its latent state being resistant to all antiviral drugs. 14, [16][17][18] Vaccines targeting this viral group have not yet been developed, which increases the need for effective therapies that not only safely treat active herpesvirus infection, but also aim to prevent latent infection.…”