学術雑誌論文

学術雑誌論文

Scientific Reports. 2017, 7(1), p. 6122

Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9

Nature Research

英語

2017-07-21

https://doi.org/10.1038/s41598-017-05905-w

Kurihara, T., Fukuhara, T., Ono, C. et al. Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9. Sci Rep 7, 6122 (2017). https://doi.org/10.1038/s41598-017-05905-w

Complete removal of hepatitis B virus (HBV) DNA from nuclei is difficult by the current therapies. Recent reports have shown that a novel genome-editing tool using Cas9 with a single-guide RNA (sgRNA) system can cleave the HBV genome in vitro and in vivo. However, induction of a double-strand break (DSB) on the targeted genome by Cas9 risks undesirable off-target cleavage on the host genome. Nickase-Cas9 cleaves a single strand of DNA, and thereby two sgRNAs are required for inducing DSBs. To avoid Cas9-induced off-target mutagenesis, we examined the effects of the expressions of nickase-Cas9 and nuclease dead Cas9 (d-Cas9) with sgRNAs on HBV replication. The expression of nickase-Cas9 with a pair of sgRNAs cleaved the target HBV genome and suppressed the viral-protein expression and HBV replication in vitro. Moreover, nickase-Cas9 with the sgRNA pair cleaved the targeted HBV genome in mouse liver. Interestingly, d-Cas9 expression with the sgRNAs also suppressed HBV replication in vitro without cleaving the HBV genome. These results suggest the possible use of nickase-Cas9 and d-Cas9 with a pair of sgRNAs for eliminating HBV DNA from the livers of chronic hepatitis B patients with low risk of undesirable off-target mutation on the host genome.

© 2017 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License.

78577

Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9

Nature Research

英語

2017-07-21

Scientific Reports

7

1

6122

https://doi.org/10.1038/s41598-017-05905-w

28733609

https://hdl.handle.net/11094/78577