Targeting Hepatitis B Virus With CRISPR/Cas9
Open accessAbstract
Hepatitis B virus persistence in infected hepatocytes is due to the presence of covalently closed circular DNA (cccDNA), the template for the transcription of viral RNAs. Antiviral therapies with nucleoside analogues inhibit replication of HBV DNA in capsids present in the cytoplasm of infected cells, but do not reduce or destroy nuclear cccDNA. To investigate whether cccDNA derived from infectious HBV could be directly targeted for destruction, we used the CRISPR/Cas9 system in HepG2 cells expressing the HBV receptor sodium taurocholate cotransporting polypeptide (NTCP). We tested different HBV-specific guide RNAs and demonstrated that they could inhibit HBV infections up to eightfold. Inhibition was due to mutations and deletions in cccDNA similar to those observed with chromosomal DNA cleaved by Cas9 and repaired by nonhomologous end joining (NHEJ). Interferon alpha (IFN-α) did not have a measurable effect on the antiviral activity of the CRISPR/Cas9 system, suggesting that Cas9 and NHEJ activities are not affected by induction of an innate immune response with the cytokine. Taken together, our results demonstrated that Cas9 can be recruited to cccDNA, opening the possibility for the development of future antiviral strategies aimed at targeting cccDNA for endonucleolytic cleavage with small molecules.
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Seeger and Sohn showed that in HepG2 cells expressing HBV, introduction of a cccDNA-targeting CRISPR-Cas9 system resulted in decreased hepatitis B core antigen expression and mutated cccDNAs.
“Recently, a study by Seeger and Sohn 28 showed that in HepG2 cells expressing HBV, the introduction of cccDNA-targeting CRISPR-Cas9 system resulted in both decreased hepatitis B core antigen expression and mutated cccDNAs”
From CRISPR-Cas9: a new and promising player in gene therapy (2015) · cites this paper as [28]
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- Authors
- Seeger, C., & Sohn, J. A.
- Affiliations
- Fox Chase Cancer Center (US)
- Corresponding author
- Seeger, Christoph
- ORCID
- Seeger, Christoph
- Venue
- Molecular Therapy - Nucleic Acids
- Publisher
- Elsevier BV
- Published
- 2014-01
- Volume
- 3
- Pages
- e216
- ISSN
- 2162-2531 (ISSN-L)
- PMC
- PMC4272409
- OpenAlex
- W2040195110
- Cited by
- 294
- References
- 44
- Citations by year
- 2026: 9 · 2025: 11 · 2024: 10 · 2023: 19 · 2022: 25 · 2021: 19 · 2020: 22 · 2019: 24 · 2018: 30 · 2017: 44 · 2016: 43 · 2015: 37
- FWCI
- 31.23
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Molecular Therapy — Nucleic Acids (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- Funders
- National Institutes of Health
- National Institute of Allergy and Infectious Diseases (R21AI101558, R21AI103514)
- Topics
- Health Sciences › Medicine › Epidemiology › Hepatitis B Virus Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Keywords
- hepatitis B virus, CRISPR/Cas9, covalently closed circular DNA, antiviral gene editing, non-homologous end joining, HepG2 cells, interferon-alpha, endonucleolytic cleavage
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