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The CRISPR/Cas9 System Facilitates Clearance of the Intrahepatic HBV Templates In Vivo

Persistence of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) under current antiviral therapy is a major barrier to eradication of chronic hepatitis B (CHB). Curing CHB will require novel strategies for specific disruption of cccDNA. The clustered regularly interspaced short palindr...

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Detalles Bibliográficos
Autores principales: Lin, Su-Ru, Yang, Hung-Chih, Kuo, Yi-Ting, Liu, Chun-Jen, Yang, Ta-Yu, Sung, Ku-Chun, Lin, You-Yu, Wang, Hurng-Yi, Wang, Chih-Chiang, Shen, Yueh-Chi, Wu, Fang-Yi, Kao, Jia-Horng, Chen, Ding-Shinn, Chen, Pei-Jer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4221598/
https://www.ncbi.nlm.nih.gov/pubmed/25137139
http://dx.doi.org/10.1038/mtna.2014.38
Descripción
Sumario:Persistence of hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) under current antiviral therapy is a major barrier to eradication of chronic hepatitis B (CHB). Curing CHB will require novel strategies for specific disruption of cccDNA. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system is a newly developed tool for site-specific cleavage of DNA targets directed by a synthetic guide RNA (gRNA) base-paired to the target DNA sequence. To examine whether this system can cleave HBV genomes, we designed eight gRNAs against HBV of genotype A. With the HBV-specific gRNAs, the CRISPR/Cas9 system significantly reduced the production of HBV core and surface proteins in Huh-7 cells transfected with an HBV-expression vector. Among eight screened gRNAs, two effective ones were identified. Interestingly, one gRNA targeting the conserved HBV sequence acted against different genotypes. Using a hydrodynamics-HBV persistence mouse model, we further demonstrated that this system could cleave the intrahepatic HBV genome-containing plasmid and facilitate its clearance in vivo, resulting in reduction of serum surface antigen levels. These data suggest that the CRISPR/Cas9 system could disrupt the HBV-expressing templates both in vitro and in vivo, indicating its potential in eradicating persistent HBV infection.