Abstract |
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.
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Authors | Su-Ru Lin, Hung-Chih Yang, Yi-Ting Kuo, Chun-Jen Liu, Ta-Yu Yang, Ku-Chun Sung, You-Yu Lin, Hurng-Yi Wang, Chih-Chiang Wang, Yueh-Chi Shen, Fang-Yi Wu, Jia-Horng Kao, Ding-Shinn Chen, Pei-Jer Chen |
Journal | Molecular therapy. Nucleic acids
(Mol Ther Nucleic Acids)
Vol. 3
Pg. e186
(Aug 19 2014)
ISSN: 2162-2531 [Print] United States |
PMID | 25137139
(Publication Type: Journal Article)
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