A knockdown of the herpes simplex virus type-1 gene in all-in-one CRISPR vectors.
Folia Histochem Cytobiol
; 58(3): 174-181, 2020.
Article
em En
| MEDLINE
| ID: mdl-32937678
ABSTRACT
INTRODUCTION:
Herpes simplex virus type 1 (HSV-1) is a virus that causes serious human disease and establishes a long-term latent infection. The latent form of this virus has shown to be resistant to antiviral drugs. Clustered Regularly Interspace Short Palindromic Repeats (CRISPR), is an important tool in genome engineering and composed of guide RNA (gRNA) and Cas9 nuclease that makes an RNA-protein complex to digest exclusive target sequences implementation of gRNA. Moreover, CRISPR-Cas9 system effectively suppresses HSV-1 infection by knockout of some viral genes. MATERIALS ANDMETHODS:
To survey the efficacy of Cas9 system on HSV-1 genome destruction, we designed several guide RNAs (gRNAs) that all packaged in one vector. Additionally, we performed a one-step restriction using BamHI and Esp3I enzymes.RESULTS:
CRISPR/Cas9 system targeted against the gD gene of HSV-1 was transfected into HEK-AD cells that showed a significant reduction of HSV-1 infection by plaque assay and real-time PCR.CONCLUSION:
The pCas-Guide-EF1a-GFP CRISPR vector can create a fast and efficient method for gRNA cloning by restriction enzymes (Esp3I (BsmBI) and BamHI). Therefore, the CRISPR/Cas9 system may be utilized for the screening of genes critical for the HSV-1 infection and developing new strategies for targeted therapy of viral infections caused by HSV-1.Palavras-chave
Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Proteínas do Envelope Viral
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Herpesvirus Humano 1
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Sistemas CRISPR-Cas
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Genes Virais
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Vetores Genéticos
Tipo de estudo:
Qualitative_research
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article