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Updated: Jun 9, 2026

Stem Cell-Derived Viral Ag-Specific T Lymphocytes Suppress HBV Replication in Mice
07:25

Stem Cell-Derived Viral Ag-Specific T Lymphocytes Suppress HBV Replication in Mice

Published on: September 25, 2019

AAV-mediated delivery of CRISPR/Cas9 targeting conserved overlapping ORFs efficiently suppresses HBV replication in

Pattida Kongsomboonchoke1, Yongyut Pewkliang2, Piyanoot Thongsri2

  • 1Department of Biotechnology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.

Biotechnology Reports (Amsterdam, Netherlands)
|June 8, 2026
PubMed

Insights

A novel CRISPR/Cas9 gene therapy delivered by adeno-associated virus (AAV) effectively targets chronic hepatitis B virus (HBV) by reducing viral DNA and cccDNA. This approach shows promise for a new generation of HBV treatments.

Area of Science:

  • Molecular Biology
  • Virology
  • Gene Therapy

Background:

  • Chronic hepatitis B virus (HBV) infection is a significant global health issue.
  • Persistent covalently closed circular DNA (cccDNA) hinders current antiviral therapies.

Purpose of the Study:

  • To develop and evaluate an adeno-associated virus (AAV)-delivered CRISPR/Cas9 system for targeting HBV.
  • To assess the efficacy of different guide RNAs (gRNAs) in reducing HBV components.

Main Methods:

  • CRISPR/Cas9 system delivered via AAV targeting conserved HBV regions.
  • Evaluation of three gRNAs (gRNA1-3) in cell lines (HepG2.2.15) and HBV-infected cells (imHCs).
  • Comparison with a reverse transcriptase-targeting gRNA and tenofovir alafenamide.

Main Results:

  • All tested gRNAs reduced intracellular and extracellular HBV DNA and HBsAg secretion.
  • gRNA2 induced a frameshift mutation, showing superior efficacy.
  • gRNA2 significantly reduced cccDNA, viral DNA, viral RNA, HBcAg, and HBsAg, with sustained suppression for 12 days.

Conclusions:

  • AAV-mediated CRISPR/Cas9 gene therapy is a promising strategy against chronic HBV infection.
  • gRNA2 demonstrated potent antiviral activity by targeting essential HBV genes.

Keywords:
AAVCRISPR/Cas9Guide RNAHBVHepatocytescccDNAimHC

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