Transient and tunable CRISPRa regulation of APOBEC/AID genes for targeting hepatitis B virus.
DNA damage repair
MT: RNA/DNA Editing
UGI
cancer
host defense
immunostimulators
machine learning
mutations
virus-host interaction
Journal
Molecular therapy. Nucleic acids
ISSN: 2162-2531
Titre abrégé: Mol Ther Nucleic Acids
Pays: United States
ID NLM: 101581621
Informations de publication
Date de publication:
13 Jun 2023
13 Jun 2023
Historique:
received:
25
10
2022
accepted:
17
04
2023
medline:
16
5
2023
pubmed:
16
5
2023
entrez:
15
5
2023
Statut:
epublish
Résumé
APOBEC/AID cytidine deaminases play an important role in innate immunity and antiviral defenses and were shown to suppress hepatitis B virus (HBV) replication by deaminating and destroying the major form of HBV genome, covalently closed circular DNA (cccDNA), without toxicity to the infected cells. However, developing anti-HBV therapeutics based on APOBEC/AID is complicated by the lack of tools for activating and controlling their expression. Here, we developed a CRISPR-activation-based approach (CRISPRa) to induce APOBEC/AID transient overexpression (>4-800,000-fold increase in mRNA levels). Using this new strategy, we were able to control APOBEC/AID expression and monitor their effects on HBV replication, mutation, and cellular toxicity. CRISPRa prominently reduced HBV replication (∼90%-99% decline of viral intermediates), deaminated and destroyed cccDNA, but induced mutagenesis in cancer-related genes. By coupling CRISPRa with attenuated sgRNA technology, we demonstrate that APOBEC/AID activation can be precisely controlled, eliminating off-site mutagenesis in virus-containing cells while preserving prominent antiviral activity. This study untangles the differences in the effects of physiologically expressed APOBEC/AID on HBV replication and cellular genome, provides insights into the molecular mechanisms of HBV cccDNA mutagenesis, repair, and degradation, and, finally, presents a strategy for a tunable control of APOBEC/AID expression and for suppressing HBV replication without toxicity.
Identifiants
pubmed: 37187708
doi: 10.1016/j.omtn.2023.04.016
pii: S2162-2531(23)00102-6
pmc: PMC10176074
doi:
Types de publication
Journal Article
Langues
eng
Pagination
478-493Informations de copyright
Crown Copyright © 2023.
Déclaration de conflit d'intérêts
The authors declare no competing interests.
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