Inhibition of replication of hepatitis B virus using transcriptional repressors that target the viral DNA.
Animals
Cell Line
CpG Islands
DNA Methylation
DNA, Circular
/ genetics
DNA, Viral
/ biosynthesis
Epigenesis, Genetic
Female
Hepatitis B
/ therapy
Hepatitis B virus
/ genetics
Liver
/ metabolism
Mice
Open Reading Frames
/ genetics
Promoter Regions, Genetic
/ genetics
Repressor Proteins
/ genetics
Virus Replication
/ genetics
DNA methylation
HBV
KRAB
TALE
Transcriptional repressor
Journal
BMC infectious diseases
ISSN: 1471-2334
Titre abrégé: BMC Infect Dis
Pays: England
ID NLM: 100968551
Informations de publication
Date de publication:
12 Sep 2019
12 Sep 2019
Historique:
received:
08
06
2019
accepted:
03
09
2019
entrez:
13
9
2019
pubmed:
13
9
2019
medline:
15
10
2019
Statut:
epublish
Résumé
Chronic infection with hepatitis B virus (HBV) is a serious global health problem. Persistence of the virus occurs as a result of stability of the replication intermediate comprising covalently closed circular DNA (cccDNA). Development of drugs that are capable of disabling this cccDNA is vital. To investigate an epigenetic approach to inactivating viral DNA, we engineered transcriptional repressors that comprise an HBV DNA-binding domain of transcription activator like effectors (TALEs) and a fused Krüppel Associated Box (KRAB). These repressor TALEs (rTALEs) targeted the viral surface open reading frame and were placed under transcription control of constitutively active or liver-specific promoters. Evaluation in cultured cells and following hydrodynamic injection of mice revealed that the rTALEs significantly inhibited production of markers of HBV replication without evidence of hepatotoxicity. Increased methylation of HBV DNA at CpG island II showed that the rTALEs caused intended epigenetic modification. Epigenetic modification of HBV DNA is a new and effective means of inactivating the virus in vivo. The approach has therapeutic potential and avoids potentially problematic unintended mutagenesis of gene editing.
Sections du résumé
BACKGROUND
BACKGROUND
Chronic infection with hepatitis B virus (HBV) is a serious global health problem. Persistence of the virus occurs as a result of stability of the replication intermediate comprising covalently closed circular DNA (cccDNA). Development of drugs that are capable of disabling this cccDNA is vital.
METHODS
METHODS
To investigate an epigenetic approach to inactivating viral DNA, we engineered transcriptional repressors that comprise an HBV DNA-binding domain of transcription activator like effectors (TALEs) and a fused Krüppel Associated Box (KRAB). These repressor TALEs (rTALEs) targeted the viral surface open reading frame and were placed under transcription control of constitutively active or liver-specific promoters.
RESULTS
RESULTS
Evaluation in cultured cells and following hydrodynamic injection of mice revealed that the rTALEs significantly inhibited production of markers of HBV replication without evidence of hepatotoxicity. Increased methylation of HBV DNA at CpG island II showed that the rTALEs caused intended epigenetic modification.
CONCLUSIONS
CONCLUSIONS
Epigenetic modification of HBV DNA is a new and effective means of inactivating the virus in vivo. The approach has therapeutic potential and avoids potentially problematic unintended mutagenesis of gene editing.
Identifiants
pubmed: 31510934
doi: 10.1186/s12879-019-4436-y
pii: 10.1186/s12879-019-4436-y
pmc: PMC6739920
doi:
Substances chimiques
DNA, Circular
0
DNA, Viral
0
Repressor Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
802Subventions
Organisme : South African National Research Foundation
ID : 81768, 81692, 68339, 85981 & 77954
Organisme : South African Poliomyelitis Research Foundation
ID : Not applicable
Organisme : German Federal Ministry for Education and Research
ID : HBVTALE-10DG15005, BMBF-01EO0803
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