https://scholars.lib.ntu.edu.tw/handle/123456789/505149
Title: | The CRISPR/Cas9 system facilitates clearance of the intrahepatic HBV templates in vivo | Authors: | Lin S.-R. HUNG-CHIH YANG Kuo Y.-T CHUN-JEN LIU Yang T.-Y Sung K.-C Lin Y.-Y HURNG-YI WANG Wang C.-C Shen Y.-C Wu F.-Y JIA-HORNG KAO DING-SHINN CHEN PEI-JER CHEN |
Issue Date: | 2014 | Publisher: | Nature Publishing Group | Journal Volume: | 3 | Start page/Pages: | e186 | Source: | Molecular Therapy - Nucleic Acids | 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 HBVexpressing templates both in vitro and in vivo, indicating its potential in eradicating persistent HBV infection. ? 2014 The American Society of Gene & Cell Therapy. All rights reserved. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-84907379292&doi=10.1038%2fmtna.2014.38&partnerID=40&md5=3e6010d556be1a0f7fe53f9519ed9f8a https://scholars.lib.ntu.edu.tw/handle/123456789/505149 |
ISSN: | 2162-2531 | DOI: | 10.1038/mtna.2014.38 | SDG/Keyword: | hepatitis B core antigen; hepatitis B surface antigen; covalently closed circular DNA; guide RNA; membrane antigen; unclassified drug; virus DNA; virus protein; animal experiment; animal model; animal tissue; antigen expression; article; controlled study; CRISPR Cas system; DNA template; expression vector; genetic transfection; genotype; hepatitis B; Hepatitis B virus genotype A; hydrodynamics; in vitro study; in vivo study; indel mutation; mouse; nonhuman; priority journal; single nucleotide polymorphism; viral clearance; virus genome; Article; clustered regularly interspaced short palindromic repeat; DNA sequence; Hepatitis B virus; human; protein expression; Southern blotting; virus genome |
Appears in Collections: | 免疫學研究所 |
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