Cargando…

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

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 for...

Descripción completa

Detalles Bibliográficos
Autores principales: Karpov, Dmitry S., Demidova, Natalia A., Kulagin, Kirill A., Shuvalova, Anastasija I., Kovalev, Maxim A., Simonov, Ruslan A., Karpov, Vadim L., Snezhkina, Anastasiya V., Kudryavtseva, Anna V., Klimova, Regina R., Kushch, Alla A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9738314/
https://www.ncbi.nlm.nih.gov/pubmed/36499174
http://dx.doi.org/10.3390/ijms232314847
_version_ 1784847509544239104
author Karpov, Dmitry S.
Demidova, Natalia A.
Kulagin, Kirill A.
Shuvalova, Anastasija I.
Kovalev, Maxim A.
Simonov, Ruslan A.
Karpov, Vadim L.
Snezhkina, Anastasiya V.
Kudryavtseva, Anna V.
Klimova, Regina R.
Kushch, Alla A.
author_facet Karpov, Dmitry S.
Demidova, Natalia A.
Kulagin, Kirill A.
Shuvalova, Anastasija I.
Kovalev, Maxim A.
Simonov, Ruslan A.
Karpov, Vadim L.
Snezhkina, Anastasiya V.
Kudryavtseva, Anna V.
Klimova, Regina R.
Kushch, Alla A.
author_sort Karpov, Dmitry S.
collection PubMed
description 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.
format Online
Article
Text
id pubmed-9738314
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-97383142022-12-11 Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems Karpov, Dmitry S. Demidova, Natalia A. Kulagin, Kirill A. Shuvalova, Anastasija I. Kovalev, Maxim A. Simonov, Ruslan A. Karpov, Vadim L. Snezhkina, Anastasiya V. Kudryavtseva, Anna V. Klimova, Regina R. Kushch, Alla A. Int J Mol Sci Article 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. MDPI 2022-11-27 /pmc/articles/PMC9738314/ /pubmed/36499174 http://dx.doi.org/10.3390/ijms232314847 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Karpov, Dmitry S.
Demidova, Natalia A.
Kulagin, Kirill A.
Shuvalova, Anastasija I.
Kovalev, Maxim A.
Simonov, Ruslan A.
Karpov, Vadim L.
Snezhkina, Anastasiya V.
Kudryavtseva, Anna V.
Klimova, Regina R.
Kushch, Alla A.
Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems
title Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems
title_full Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems
title_fullStr Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems
title_full_unstemmed Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems
title_short Complete and Prolonged Inhibition of Herpes Simplex Virus Type 1 Infection In Vitro by CRISPR/Cas9 and CRISPR/CasX Systems
title_sort complete and prolonged inhibition of herpes simplex virus type 1 infection in vitro by crispr/cas9 and crispr/casx systems
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9738314/
https://www.ncbi.nlm.nih.gov/pubmed/36499174
http://dx.doi.org/10.3390/ijms232314847
work_keys_str_mv AT karpovdmitrys completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT demidovanataliaa completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT kulaginkirilla completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT shuvalovaanastasijai completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT kovalevmaxima completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT simonovruslana completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT karpovvadiml completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT snezhkinaanastasiyav completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT kudryavtsevaannav completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT klimovareginar completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems
AT kushchallaa completeandprolongedinhibitionofherpessimplexvirustype1infectioninvitrobycrisprcas9andcrisprcasxsystems