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Targeting hepatitis B virus cccDNA by CRISPR/Cas9 nuclease efficiently inhibits viral replication
Targeting hepatitis B virus cccDNA by CRISPR/Cas9 nuclease efficiently inhibits viral replication
- Source :
- Antiviral research. 118
- Publication Year :
- 2014
-
Abstract
- Chronic hepatitis B virus (HBV) infection causes liver cirrhosis and hepatocellular carcinoma and remains a serious health problem worldwide. Covalently closed circular DNA (cccDNA) in the liver cell nucleus sustains HBV infection. Major treatments for HBV infection include the use of interferon-α and nucleotide analogs, but they cannot eradicate cccDNA. As a novel tool for genome editing, clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system developed from bacteria can be used to accurately and efficiently engineer and modify genomic DNA. In this study, the CRISPR/Cas9 system was used to target the HBV genome and efficiently inhibit HBV infection. We synthesized four single-guide RNAs (sgRNAs) targeting the conserved regions of HBV. The expression of these sgRNAS with Cas9 reduced the viral production in Huh7 cells as well as in HBV-replication cell HepG2.2.15. We further demonstrated that CRISPR/Cas9 direct cleavage and cleavage-mediated mutagenesis occurred in HBV cccDNA of transfected cells. In the new mouse model carrying HBV cccDNA, injection of sgRNA–Cas9 plasmids via rapid tail vein resulted in the low level of cccDNA and HBV protein. In conclusion, the designed CRISPR/Cas9 system can accurately and efficiently target HBV cccDNA and inhibit HBV replication. This system may be used as a novel therapeutic strategy against chronic HBV infection.
- Subjects :
- HBV RNA encapsidation signal epsilon
Hepatitis B virus
CRISPR-Associated Proteins
Biology
medicine.disease_cause
Virus Replication
Antiviral Agents
Virus
Cell Line
Hepatitis B, Chronic
Virology
medicine
CRISPR
Animals
Humans
Pharmacology
Drug Carriers
Mice, Inbred BALB C
Cas9
Liver cell
virus diseases
cccDNA
Endonucleases
Molecular biology
digestive system diseases
Disease Models, Animal
Treatment Outcome
Viral replication
DNA, Viral
Hepatocytes
Female
DNA, Circular
Plasmids
Subjects
Details
- ISSN :
- 18729096
- Volume :
- 118
- Database :
- OpenAIRE
- Journal :
- Antiviral research
- Accession number :
- edsair.doi.dedup.....38f50f7b7489f6447e88abff754d38b5