Suppressing the NHEJ pathway by DNA-PKcs inhibitor NU7026 prevents degradation of HBV cccDNA cleaved by CRISPR/Cas9.
Apoptosis
CRISPR-Cas Systems
Catalytic Domain
Cell Survival
Chromones
/ pharmacology
DNA End-Joining Repair
/ drug effects
DNA Mutational Analysis
DNA, Circular
/ drug effects
DNA, Viral
/ drug effects
Gene Deletion
Genome, Viral
Hep G2 Cells
Hepatitis B virus
Hepatitis B, Chronic
/ metabolism
High-Throughput Nucleotide Sequencing
Humans
Morpholines
/ pharmacology
RNA, Guide, Kinetoplastida
/ genetics
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
12 02 2019
12 02 2019
Historique:
received:
08
08
2018
accepted:
31
12
2018
entrez:
14
2
2019
pubmed:
14
2
2019
medline:
24
9
2020
Statut:
epublish
Résumé
Chronic hepatitis B is a severe liver disease caused by hepatitis B virus (HBV) infection. Covalently closed circular DNA (cccDNA), a super-spiralized, double-stranded form of the HBV genome, is the major determinant of viral persistence. CRISPR/Cas9 nucleases have been recently shown to introduce double-stranded DNA breaks into HBV cccDNA. The inflicted damage results predominantly in erroneous repair of cccDNA by non-homologous end-joining (NHEJ). NHEJ has been suggested to enhance anti-HBV activity of CRISPR/Cas9 and increase cccDNA mutation. In this study, we assessed anti-HBV activity of CRISPR/Cas9 and cccDNA repair outcomes in an altered NHEJ/HR environment. NU7026, a strong inhibitor of NHEJ, prevented CRISPR/Cas9-mediated degradation of cccDNA and resulted in frequent on-target deletions. We conclude that CRISPR/Cas9 is a highly effective tool to degrade cccDNA and first demonstrate that inhibiting NHEJ impairs cccDNA degradation.
Identifiants
pubmed: 30755668
doi: 10.1038/s41598-019-38526-6
pii: 10.1038/s41598-019-38526-6
pmc: PMC6372644
doi:
Substances chimiques
2-(morpholin-4-yl)benzo(h)chromen-4-one
0
Chromones
0
DNA, Circular
0
DNA, Viral
0
Morpholines
0
RNA, Guide
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
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