ライフサイエンスコーパス: cccDNA

1 cccDNA is assembled with cellular histone proteins into

2 cccDNA is maintained in the nucleus of the infected hepa

3 cccDNA transcription is reduced in HIV/HBV coinfected pe

4 he nuclei assayed contained between 1 and 17 cccDNA molecules, with the remaining 10% containing more

5 ssing the virus are known, information about cccDNA formation, stability, and turnover is lacking.

6 Current knowledge about cccDNA stability was largely derived from quantitative a

7 T1 and STAT2 transcription factors to active cccDNA.

8 the enhancer I HBV regulatory region altered cccDNA transcription and viral replication.

9 relationship between the UV-DDB complex and cccDNA formation, providing a proof of concept for a mor

10 he relationship between hepatocyte death and cccDNA elimination requires knowing both the amount of h

11 cular mechanisms of rcDNA deproteination and cccDNA biogenesis.IMPORTANCE The covalently closed circu

12 nderstanding of HBV rcDNA deproteination and cccDNA biosynthesis.

13 assay for the detection of HBV RNA, DNA, and cccDNA.

14 en-positive (HBsA-positive) and HBV DNA- and cccDNA-positive cells.

15 c cccDNA but not between HBeAg or HBcrAg and cccDNA in HBV-infected chimpanzees, suggesting that HBc

16 oplasm and reduction of nuclear DP rcDNA and cccDNA.

17 ate HBV genomic reservoirs, which persist as cccDNA.

18 3.5 kB HBV RNA (preC/pgRNA) and by assessing cccDNA-associated histone tails post-transcriptional mod

19 d the decline in cccDNA, so that the average cccDNA copy number in infected cells dropped during the

20 Because cccDNA does not replicate semiconservatively, it is not

21 A reduced association between cccDNA and P300 (mean % input: 0.69-1.81% vs. 3.48% in w

22 iated cccDNA targeting were compared between cccDNAs with distinct transcriptional activities.

23 , and the reduction rate was similar between cccDNAs in transcriptionally active and transcriptionall

24 DNA by Cas9 and a dramatic reduction in both cccDNA and other parameters of viral gene expression and

25 ear episomal form of the viral genome called cccDNA, the most stable HBV replication intermediate.

26 motes gene expression from the chromatinized cccDNA, accompanied by an epigenetic switch from an H3K9

27 of intrahepatic covalently closed circular (cccDNA), which is an important target for pegylated inte

28 To fill the gaps in knowledge concerning cccDNA biology, we have developed a fluorescence imaging

29 In contrast, cccDNA declined more slowly, consistent with a half-life

30 essed by qPCR, RCA and ddPCR assays detected cccDNA in all-but-one negative samples.

31 gating HBV cccDNA biology and for developing cccDNA-targeting drugs.

32 ded the first insights into the fate of DHBV cccDNA and nuclear HBV DNA under conditions mimicking an

33 e detection of duck hepatitis B virus (DHBV) cccDNA and HBV nuclear DNA in established cell lines.

34 -polymerase inhibitor aphidicolin diminishes cccDNA formation both in biochemical assays and in HBV-i

35 atin remodelers for their ability to disrupt cccDNA biosynthesis and function.

36 strated that these inhibitors also disrupted cccDNA synthesis during de novo HBV infection of HepG2 c

37 blishment of covalently closed circular DNA (cccDNA) and can be either transient or persistent.

38 s, including covalently closed circular DNA (cccDNA) and Dane particles, were detected only after ind

39 ids produced covalently closed circular DNA (cccDNA) and HBV early antigen (HBeAg), expressed intrace

40 an episomal covalently-closed-circular DNA (cccDNA) and integrated viral DNA fragments are linked to

41 (HBsAg) from covalently closed circular DNA (cccDNA) and the integrated genome, is believed to contri

42 e same time, covalently closed circular DNA (cccDNA) and viral mRNA levels both declined about two- t

43 es including covalently closed circular DNA (cccDNA) are present.

44 fect initial covalently closed circular DNA (cccDNA) conversion but inhibits the synthesis of progeny

45 f the stable covalently closed circular DNA (cccDNA) form of the viral genome, which is formed by the

46 coating, and covalently closed circular DNA (cccDNA) formation.

47 Covalently closed circular DNA (cccDNA) forms the basis for replication and persistence

48 of the viral covalently closed circular DNA (cccDNA) genome and host histones.

49 s methylated covalently closed circular DNA (cccDNA) in human liver tissue.

50 umulation of covalently closed circular DNA (cccDNA) in nuclei of infected cells.

51 erted into a covalently closed circular DNA (cccDNA) in the host cell nucleus.

52 lear pool of covalently closed circular DNA (cccDNA) in the liver.

53 d by the persistence of closed circular DNA (cccDNA) in the nucleus of infected hepatocytes.

54 erted into a covalently closed circular DNA (cccDNA) in the nucleus of the infected hepatocyte by cel

55 HBV covalently closed circular DNA (cccDNA) is a major obstacle for a cure of chronic hepati

56 virus (HBV) covalently closed circular DNA (cccDNA) is an episomal minichromosome whose persistence

57 PORTANCE The covalently closed circular DNA (cccDNA) is the persistent form of the hepatitis B virus

58 from the HBV covalently closed circular DNA (cccDNA) minichromosome, both in cultured cells in which

59 a long-lived covalently closed circular DNA (cccDNA) molecule, is degraded noncytolytically by agents

60 V) contained covalently closed circular DNA (cccDNA) molecules with deletions and insertions indicati

61 Covalently closed circular DNA (cccDNA) of hepadnaviruses exists as an episomal minichro

62 Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is formed by conversi

63 HBV covalently closed circular DNA (cccDNA) plays an essential role in HBV persistence and r

64 virus (HBV) covalently closed circular DNA (cccDNA) requires the removal of the covalently linked vi

65 ool of viral covalently closed circular DNA (cccDNA) transcriptional template of HBV, which must be e

66 ated DNA and covalently closed circular DNA (cccDNA) were turned over independently of cell division.

67 virus (HBV), covalently closed circular DNA (cccDNA), has been difficult to study in patients with ch

68 rsistence of covalently closed circular DNA (cccDNA), HBV-DNA integration into the host genome, and i

69 episome, the covalently closed circular DNA (cccDNA), in hepatocytes.

70 episome, the covalently closed circular DNA (cccDNA), in the nuclei of infected hepatocytes.

71 omosome, the covalently closed circular DNA (cccDNA), in the nucleus of infected hepatocytes, as well

72 al template, covalently closed circular DNA (cccDNA), is long lived in infected hepatocytes.

73 ool of viral covalently closed circular DNA (cccDNA), resulting in a transient elevation of viral rep

74 ctly affect nuclear HBV closed circular DNA (cccDNA), the genomic form that templates viral transcrip

75 formation of covalently closed circular DNA (cccDNA), the molecular basis for establishing and sustai

76 Covalently closed circular DNA (cccDNA), the nuclear form of hepatitis B virus (HBV), is

77 ation of new covalently closed circular DNA (cccDNA), the viral transcriptional template.

78 silencing of covalently closed circular DNA (cccDNA), to characterize this dissociation, and virologi

79 te the viral covalently closed circular DNA (cccDNA), which is a stable episomal form of the viral ge

80 e stable HBV covalently-closed-circular DNA (cccDNA).

81 ncluding the covalently closed circular DNA (cccDNA).

82 hepadnavirus covalently closed circular DNA (cccDNA).

83 ahepatic WHV covalently closed circular DNA (cccDNA).

84 HBV covalently closed episomal circular DNA (cccDNA).

85 ion was from covalently closed circular DNA (cccDNA).

86 forms into a covalently closed circular DNA (cccDNA).

87 genome, the covalently closed circular DNA (cccDNA).

88 ral genomes [covalently closed circular DNA (cccDNA)] in the nuclei of infected cells.

89 ue to the persistence of viral episomal DNA (cccDNA) in infected cells.

90 ch resembles covalently closed circular DNA [cccDNA]).

91 transcriptase activity but fail to eliminate cccDNA, which would be required to cure HBV infection.

92 proposed as surrogate markers for evaluating cccDNA activity, they do not necessarily estimate the am

93 It remains unclear if existing cccDNA is eliminated noncytolytically or if hepatocyte d

94 The development of a FISH-based assay for cccDNA tracking provided the first insights into the fat

95 d samples was associated with enrichment for cccDNA histone PTMs related to repressed transcription.

96 of lagging-strand synthesis as essential for cccDNA formation: proliferating cell nuclear antigen, th

97 ishing these as a minimal set of factors for cccDNA formation.

98 roteinated-rcDNA (DP-rcDNA) intermediate for cccDNA formation.

99 While methods for cccDNA quantification from liver biopsy specimens and ce

100 Each nucleus was assayed by nested PCR for cccDNA and for cellular IFN-alpha genes as an internal c

101 ntification of key host factors required for cccDNA metabolism and function will reveal molecular tar

102 circular-DNA substrates are repaired to form cccDNA by both cell extracts and purified human proteins

103 cer and the amount of pgRNA transcribed from cccDNA) were significantly higher in cells expressing wi

104 ISGs) on hepadnaviral mRNAs transcribed from cccDNA, we found that downregulating the expression of S

105 acterize residual HBV-RNA transcription from cccDNA and assist drug development and disease managemen

106 acterize residual HBV RNA transcription from cccDNA, and assist new drug development and disease mana

107 The mcHBV-GLuc cccDNA model is independent of HBV infection, and will b

108 d to non-HBV minicircle plasmids, mcHBV-GLuc cccDNA showed persistent HBV-GLuc activity and HBx-depen

109 Importantly, the mcHBV-GLuc cccDNA showed resistance to interferons (IFN) treatment,

110 The mcHBV-GLuc cccDNA was easily produced in bacteria, and it formed mi

111 th a Gaussia Luciferase reporter (mcHBV-GLuc cccDNA), which serves as a surrogate to measure cccDNA a

112 HBV cccDNA is converted from viral genomic relaxed circular

113 HBV cccDNA levels remained stable in nondividing hepatocytes

114 HBV cccDNA minichromosomes in distinct epigenetic transcript

115 cteria, and it formed minichromosomes as HBV cccDNA episome DNA does when it was transfected into hum

116 and nucleic acids and a mismatch between HBV cccDNA, RNA, and expression of the hepatitis B surface a

117 B virus covalently closed circular DNA (HBV cccDNA), the episomal form of the virus that persists de

118 es and epigenetic modifiers specific for HBV cccDNA.

119 only, host DNA-repair factor involved in HBV cccDNA biogenesis.

120 r, our findings define key components in HBV cccDNA formation.

121 V entry inhibitors, HBV core inhibitors, HBV cccDNA transcripts RNA interference, HBV cell apoptosis

122 mechanisms, we quantitated intrahepatic HBV cccDNA levels in acutely infected chimpanzees whose viro

123 , and will be valuable for investigating HBV cccDNA biology and for developing cccDNA-targeting drugs

124 We engineered a minicircle HBV cccDNA with a Gaussia Luciferase reporter (mcHBV-GLuc cc

125 We report here a novel HBV cccDNA technology that will meet the need.

126 HBVcircle models, the reduction rate of HBV cccDNA and the efficacy of apolipoprotein B mRNA editing

127 a, produced by T cells, reduce levels of HBV cccDNA in hepatocytes by inducing deamination and subseq

128 n about the intrahepatic distribution of HBV cccDNA in infected patients, particularly at the single-

129 nistic understanding of the stability of HBV cccDNA in the presence of antiviral therapy and during c

130 Elimination of HBV cccDNA is so far only achieved by antiviral immune respo

131 I and II may catalyze distinct steps of HBV cccDNA synthesis and that pharmacologic targeting of the

132 -alpha mediates epigenetic repression of HBV cccDNA transcriptional activity, which may assist in the

133 cccDNA is hampered by the lack of robust HBV cccDNA models.

134 to treat HBV do not directly target the HBV cccDNA genome and thus cannot eradicate HBV infection.

135 cal host-protein interaction site in the HBV cccDNA genome may facilitate the development of novel an

136 ent, indicating its unique similarity to HBV cccDNA that is usually resistant to long-term IFN treatm

137 el it also decreased hepatitis B virus (HBV) cccDNA and woodchuck surface antigen.

138 55-60 and 121-126 had a lower degree of HBx-cccDNA association than wild type HBx (mean % input: 0.0

139 mediate IFN-alpha silencing of hepadnaviral cccDNA transcription.

140 pressive effect of IFN-alpha on hepadnaviral cccDNA transcription.IMPORTANCE Pegylated IFN-alpha is t

141 Most importantly, GLuc illuminates cccDNA as a surrogate of cccDNA activity, providing a ve

142 s revealed differential promoter activity in cccDNA and integrants, with implications for the efficac

143 ted hepatocytes lagged behind the decline in cccDNA, so that the average cccDNA copy number in infect

144 Moreover, the declines in cccDNA appeared to reflect the loss of hepatocytes from

145 ith M and/or S proteins led to a decrease in cccDNA levels, indicating that L contributes to the regu

146 No difference in cccDNA levels was found according to serum viral markers

147 and minimal set of host factors involved in cccDNA formation is unknown, largely due to the lack of

148 tform, we found that nucleosome occupancy in cccDNA regulates X transcription.

149 ledge, the first genome-wide maps of PTMs in cccDNA-containing chromatin from de novo infected HepG2

150 gate the role of the cytoplasmic DP rcDNA in cccDNA formation, we demonstrated that rcDNA deproteiniz

151 Administration of IFN-alpha resulted in cccDNA-bound histone hypoacetylation as well as active r

152 t with TPD2's having a physiological role in cccDNA formation, RNAi-mediated TDP2 depletion in human

153 s their inability to eradicate or inactivate cccDNA.

154 for detecting viral nucleic acids, including cccDNA, with single-cell resolution provides a means for

155 Initially, cccDNA is derived from RC DNA from the infecting virion,

156 r DNA (rcDNA) and subsequent conversion into cccDNA within 12 to 24 h.

157 A formation, and conversion of DP-rcDNA into cccDNA is a rate-limiting step of cccDNA formation in He

158 ation was found between HBc and intrahepatic cccDNA but not between HBeAg or HBcrAg and cccDNA in HBV

159 the increase of HBV viremia and intrahepatic cccDNA loads was significantly slower than in HBV mono-i

160 HBc can be a better marker for intrahepatic cccDNA.

161 Clearances of intrahepatic cccDNA and/or HBsAg are critical endpoints for future an

162 cted the amount and dynamics of intrahepatic cccDNA within a certain range using specific viral marke

163 were used to quantify residual intrahepatic cccDNA in liver biopsies from 56 chronically HBV infecte

164 The assessment of residual intrahepatic cccDNA levels and activity after long-term nucleos(t)ide

165 DNA), which serves as a surrogate to measure cccDNA activity.

166 We have therefore measured cccDNA levels in woodchuck hepatocyte cultures following

167 d CRISPR/CRISPR-associated 9 (Cas9)-mediated cccDNA targeting were compared between cccDNAs with dist

168 Furthermore, methylated cccDNA was found in tumor and nonneoplastic human liver

169 Here, using a new cccDNA ChIP-Seq approach, we report, to our knowledge, t

170 actions that can be exploited to develop new cccDNA-targeting antivirals.

171 he generation time of hepatocytes and if new cccDNA formation were effectively blocked.

172 is possible to segregate cells containing no cccDNA.

173 K7A and K7R mutations did not affect de novo cccDNA formation and RNA transcription during infection,

174 The nuclear cccDNA pool can be replenished through recycling of newl

175 ons regulate the transcriptional activity of cccDNA minichromosomes.

176 up to 36 days with 3TC reduced the amount of cccDNA in the cultures not more than twofold compared to

177 e and quick method to detect trace amount of cccDNA.

178 ey do not necessarily estimate the amount of cccDNA.

179 in limits the intracellular amplification of cccDNA for duck hepatitis B virus.

180 proteins in regulating the amplification of cccDNA in HBV is not well characterized.

181 synthesis and intracellular amplification of cccDNA.

182 argely derived from quantitative analyses of cccDNA levels present in liver samples, and little was k

183 hosen guide RNAs, we demonstrate cleavage of cccDNA by Cas9 and a dramatic reduction in both cccDNA a

184 e infecting virion, but additional copies of cccDNA are derived from newly synthesized RC DNA molecul

185 ect effect on the up to 50 or more copies of cccDNA that maintain the infected state.

186 ma and TNF-alpha each induced deamination of cccDNA and interfered with its stability; their effects

187 Deprivation of cccDNA required activation of nuclear APOBEC3 deaminases

188 In vitro, the desiccation of cccDNA led to loss of supercoiling, aggregation, loss of

189 sing FISH, we determined the distribution of cccDNA under conditions mimicking chronic infections wit

190 The underlying dynamics of cccDNA persistence are unlikely to impact the probabilit

191 te death were responsible for elimination of cccDNA during recovery from transient infections.

192 However, elimination of cccDNA might be facilitated if its half-life were short

193 s one of the major routes for elimination of cccDNA.

194 Finally, an in vitro equivalent of cccDNA showed decreased viral protein production in HepG

195 articular, little is known about the fate of cccDNA during cell division.

196 ples, and little was known about the fate of cccDNA in individual cells.

197 t with the possibility that some fraction of cccDNA was distributed to daughter cells in those infect

198 the formation of two or more generations of cccDNA from linear DNA was observed.

199 ing of antiviral target and investigation of cccDNA biosynthesis and transcription using secreted HBs

200 DNA target will promote the highest level of cccDNA disruption.

201 kemia (PML) protein increased basal level of cccDNA transcription activity and partially attenuated I

202 ion led to a decrease (>50%) in the level of cccDNA, which inversely correlated with the level of the

203 led to an increase (>6-fold) in the level of cccDNA.

204 of viral RNAs without altering the level of cccDNA.

205 ation, the cells accumulate higher levels of cccDNA as well as larger amounts of deproteinized rcDNA

206 hibitors significantly reduced the levels of cccDNA.

207 The loss of cccDNA was comparable to that expected from the estimate

208 ring antiviral therapy, the rates of loss of cccDNA, infected hepatocytes (1 or more molecules of ccc

209 ter T-cell stimulation prevented the loss of cccDNA.

210 tial for the biosynthesis and maintenance of cccDNA and reveal that cellular DNA topoisomerases are r

211 The mechanism of cccDNA formation is unknown, but the release of P protei

212 ce suggests that epigenetic modifications of cccDNA contribute to viral replication and the outcome o

213 nduce the posttranslational modifications of cccDNA-associated histones similar to those induced by I

214 a-induced posttranslational modifications of cccDNA-associated histones.

215 infected hepatocytes (1 or more molecules of cccDNA), and replicating DNAs may be quite different.

216 tudies have measured the mean copy number of cccDNA molecules in hepadnaviral-infected cells, the dis

217 a PCR-based assay, we examined the number of cccDNA molecules of the duck hepatitis B virus in single

218 all, the data suggest (i) that the number of cccDNA molecules per cell may fluctuate over time, and (

219 This panel of cccDNA evaluation techniques should provide an added val

220 To study the molecular pathway of cccDNA formation and its regulation by viral and cellula

221 ormed to determine the epigenetic pattern of cccDNA.

222 The first step reduces the pool of cccDNA molecules noncytolytically, probably by eliminati

223 n, which is associated with the reduction of cccDNA-associated histone modifications specifying activ

224 e, including the mechanism and regulation of cccDNA formation.

225 ssibility that chromatin-based regulation of cccDNA transcription could be a new therapeutic approach

226 ting that L contributes to the regulation of cccDNA.

227 Epigenetic sensitization of cccDNA makes it more susceptible to damage and may poten

228 RNA and HBcrAg as surrogates of silencing of cccDNA, to characterise this dissociation, and virologic

229 rcDNA into cccDNA is a rate-limiting step of cccDNA formation in HepG2 cells.

230 lar proteins that mediate the suppression of cccDNA transcription by the cytokine, we found that down

231 artially attenuated IFN-alpha suppression of cccDNA transcription.

232 y, GLuc illuminates cccDNA as a surrogate of cccDNA activity, providing a very sensitive and quick me

233 port demonstrates regulation of synthesis of cccDNA by the envelope proteins of HBV.

234 hepatocytes from the cultures rather than of cccDNA from hepatocytes.

235 espite this multipronged response, traces of cccDNA persist indefinitely in the liver, likely providi

236 ive biomarkers of continued transcription of cccDNA in HBeAg-negative patients despite marked HBV DNA

237 ive biomarkers of continued transcription of cccDNA in HBeAg-negative patients despite marked HBV-DNA

238 A fraction of cccDNAs formed from both type 1 and type 2 linear DNAs a

239 cular inhibitors of DNA metabolic enzymes on cccDNA synthesis but avoids cytotoxic effects upon long-

240 tion and spread but have no direct impact on cccDNA or expression of viral genes.

241 HBV replication with little or no impact on cccDNA, hence lifelong treatment is required in the vast

242 not markedly affect rcDNA deproteination or cccDNA formation.

243 tion products, without affecting HBV RNAs or cccDNA.

244 The specificity of our cccDNA probe set was confirmed by its strict intranuclea

245 The amount of pgRNA transcribed per cccDNA also decreased from HB1 to HB5 (P < .05).

246 ersion but inhibits the synthesis of progeny cccDNA by amplification.

247 exposure to >7 years exposure), quantifying cccDNA and pregenomic RNA (pgRNA) in each cell using dro

248 ased assay supporting synchronized and rapid cccDNA synthesis from intracellular progeny nucleocapsid

249 models and in vitro and in vivo recombinant cccDNA (rcccDNA) and HBVcircle models, the reduction rat

250 We reconstituted cccDNA formation with purified human homologues, establi

251 biochemical system that fully reconstitutes cccDNA formation.

252 alpha, inhibited HBV replication and reduced cccDNA in infected cells without the direct contact requ

253 n which shRNA-DDB1 in HepDES19 cells reduced cccDNA production.

254 wn of topoisomerase II significantly reduced cccDNA amplification.

255 STAT1, SMCHD1, or PML significantly reduced cccDNA transcription activity.

256 at HBV envelope proteins negatively regulate cccDNA formation, and conversion of DP-rcDNA into cccDNA

257 rs, inhibitors of subviral particle release, cccDNA silencers, and RNA interference molecules have re

258 e infected hepatocytes, are needed to remove cccDNA from surviving infected hepatocytes.

259 Activity of residual cccDNA was evaluated by quantifying 3.5 kB HBV RNA (preC

260 anti-HBV therapeutics against the resilient cccDNA template.

261 n of novel targets to destabilize or silence cccDNA.

262 umanized livers, suggesting that spontaneous cccDNA clearance may occur.

263 cytes by inducing deamination and subsequent cccDNA decay.

264 lpha-induced cellular proteins that suppress cccDNA transcription and may partly mediate IFN-alpha si

265 he development of drugs that directly target cccDNA is hampered by the lack of robust HBV cccDNA mode

266 Novel approaches that directly target cccDNA regulation would therefore be highly desirable.

267 at replicative DNA levels declined more than cccDNA and mRNA levels following adenovirus infection su

268 and functional studies, we demonstrate that cccDNA indeed contains ten G4s structures.

269 se observations, our study demonstrated that cccDNA was undetectable in a portion of HBV-infected cel

270 Collectively, ddPCR revealed that cccDNA content is stable during hepatocyte proliferation

271 Collectively, these results suggest that cccDNA clearance is a two-step process mediated by the c

272 Finally, we showed for the first time that cccDNA undergoes phase separation in a G4-dependent mann

273 iral DNA found in the cytoplasm but also the cccDNA from the nucleus.

274 therapeutic anti-HBV agents, eliminating the cccDNA remains challenging.

275 nstrate that the elimination kinetics of the cccDNA are more rapid than the elimination of HBV antige

276 In contrast, terminal clearance of the cccDNA is associated with the peak of liver disease and

277 ssion decreased rapidly-less than 50% of the cccDNA remained detectable in 1.5 days.

278 and its administration timing to reduce the cccDNA counts.

279 ylation as well as active recruitment to the cccDNA of transcriptional corepressors.

280 This cccDNA formation assay allows systematic screening of th

281 P2 inhibitors reduced the production of this cccDNA synthesis intermediate to a lesser extent.

282 ications (PTMs) of histone proteins bound to cccDNA through analysis of de novo HBV-infected cell lin

283 ificantly slowed the conversion of RC-DNA to cccDNA.

284 recruitment of histone modifying enzymes to cccDNA.

285 hat those cellular proteins are recruited to cccDNA minichromosomes and induce the posttranslational

286 hat STAT1, SMCHD1, and PML were recruited to cccDNA minichromosomes and phenocopied the IFN-alpha-ind

287 st, enhanced expression of S RNA relative to cccDNA and core RNA in patients with medium-high or low

288 plication, antigen expression, and, in turn, cccDNA activity, providing a potential path to a functio

289 spite 38 out of 56 patients had undetectable cccDNA when assessed by qPCR, RCA and ddPCR assays detec

290 Upper cccDNA range and median pgRNA decreased from HB1 to HB5

291 hat maintenance of wild-type levels of viral cccDNA promotes persistence of virus infection by establ

292 ed birds coincided with a reduction of viral cccDNA to wild-type virus levels in the liver.

293 aimed at eliminating or crippling the viral cccDNA.

294 Hepatitis B virus cccDNA is the key target for the necessary development o

295 pression of human and duck hepatitis B virus cccDNA transcription, which is associated with the reduc

296 ultiple enzymes which target different vital cccDNA regions, or sequential delivery of different enzy

297 life cycle, little is understood about where cccDNA molecules localize, why they are so stable, and h

298 he amount of hepatocyte turnover and whether cccDNA synthesis is effectively blocked during the perio

299 analysis of the recombination joints in WHV cccDNA.

300 gulation, via their impeded interaction with cccDNA and altered recruitment of histone modifying enzy