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Generating mRNA Encoding Anti-HBV Designer Epigenome Modifiers.

Prashika Singh1, Tafadzwa Mlambo2, Kristie Bloom1

  • 1Wits/SAMRC Antiviral Gene Therapy Research Unit, Infectious Diseases and Oncology Research Institute (IDORI), Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.

Methods in Molecular Biology (Clifton, N.J.)
|August 28, 2025
PubMed
Summary
This summary is machine-generated.

New epigenome editing tools use designer epigenome modifiers (DEMs) delivered via mRNA to target and silence chronic hepatitis B virus (HBV) replication. This approach offers a promising, non-mutagenic strategy for lasting HBV therapy.

Keywords:
CpG islandsDEMsHBVIn vitro transcription

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Area of Science:

  • Epigenetics
  • Virology
  • Molecular Biology

Background:

  • Chronic hepatitis B virus (HBV) infection affects over 257 million people globally, causing significant mortality from liver disease.
  • Current antiviral therapies are limited in their ability to eliminate the persistent HBV covalently closed circular DNA (cccDNA) reservoir.
  • Epigenome editing presents a novel, non-mutagenic strategy to target viral gene expression.

Purpose of the Study:

  • To develop and evaluate mRNA-encoded designer epigenome modifiers (DEMs) for targeted epigenetic silencing of HBV.
  • To optimize mRNA production by removing immunostimulatory double-stranded RNA (dsRNA).
  • To assess the efficacy of DEM-mediated transcriptional repression of HBV DNA in cell culture.

Main Methods:

  • Production of mRNA encoding DEMs engineered to methylate CpG islands in key HBV open reading frames (ORFs).
  • In vitro transcription of DEM sequences from the pT7(AG) platform.
  • Purification of mRNA to remove dsRNA contaminants, enhancing expression.
  • Transfection of cultured cells with synthesized mRNA and assessment of HBV DNA transcriptional repression.

Main Results:

  • Successful synthesis of mRNA encoding DEMs designed for HBV epigenome modification.
  • Improved mRNA expression achieved through dsRNA removal protocols.
  • Demonstrated targeted transcriptional repression of HBV DNA in cultured cells mediated by DEMs.

Conclusions:

  • mRNA-delivered DEMs represent a viable strategy for epigenetic silencing of chronic HBV.
  • This approach minimizes risks of genomic integration and off-target effects associated with other gene-editing technologies.
  • Epigenetic therapy holds potential for durable therapeutic benefits in chronic HBV infection.