CRISPR/Cas9 nickase-mediated disruption of hepatitis B virus open reading frame S and X

被引：106

作者：

Karimova, Madina ^{[1
]}

Beschorner, Niklas ^{[1
]}

Dammermann, Werner ^{[2
]}

Chemnitz, Jan ^{[1
]}

Indenbirken, Daniela ^{[1
]}

Bockmann, Jan-Hendrik ^{[2
]}

Grundhoff, Adam ^{[1
,3
]}

Lueth, Stefan ^{[2
]}

Buchholz, Frank ^{[4
,5
]}

zur Wiesch, Julian Schulze ^{[2
,3
]}

Hauber, Joachim ^{[1
,3
]}

机构：

[1] Leibniz Inst Expt Virol, Heinrich Pette Inst, D-20251 Hamburg, Germany

[2] Univ Med Ctr Eppendorf, Dept Med 1, D-20246 Hamburg, Germany

[3] German Ctr Infect Res DZIF, Hamburg, Germany

[4] Univ Hosp, Dept Med Syst Biol, D-01307 Dresden, Germany

[5] Tech Univ Dresden, Med Fac Carl Gustav Carus, D-01307 Dresden, Germany

来源：

SCIENTIFIC REPORTS | 2015年 / 5卷

关键词：

CLOSED CIRCULAR DNA; CELLS; CCCDNA; CAS9; GENE; TRANSCRIPTION; CRISPR-CAS9; CLEAVAGE; REPLICATION; STRATEGIES;

D O I：

10.1038/srep13734

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Current antiviral therapies cannot cure hepatitis B virus (HBV) infection; successful HBV eradication would require inactivation of the viral genome, which primarily persists in host cells as episomal covalently closed circular DNA (cccDNA) and, to a lesser extent, as chromosomally integrated sequences. However, novel designer enzymes, such as the CRISPR/Cas9 RNA-guided nuclease system, provide technologies for developing advanced therapy strategies that could directly attack the HBV genome. For therapeutic application in humans, such designer nucleases should recognize various HBV genotypes and cause minimal off-target effects. Here, we identified cross-genotype conserved HBV sequences in the S and X region of the HBV genome that were targeted for specific and effective cleavage by a Cas9 nickase. This approach disrupted not only episomal cccDNA and chromosomally integrated HBV target sites in reporter cell lines, but also HBV replication in chronically and de novo infected hepatoma cell lines. Our data demonstrate the feasibility of using the CRISPR/Cas9 nickase system for novel therapy strategies aiming to cure HBV infection.

引用

页数：16

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