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CRISPR/Cas9 nickase-mediated disruption of hepatitis B virus open reading frame S and X
Madina Karimova, Niklas Beschorner, Werner Dammermann, Jan Chemnitz, Daniela Indenbirken, Jan-Hendrik Bockmann, Adam Grundhoff, Stefan Lüth, Frank Buchholz, Julian Schulze zur Wiesch & Joachim Hauber
Scientific Reports 5, Article number: 13734 (2015)
doi:10.1038/srep13734
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Cas9 endonuclease | Hepatitis B
Received:
11 January 2015
Accepted:
04 August 2015
Published online:
03 September 2015
Abstract
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.
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发表于 2015-9-11 15:43
