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

Takeshi Kurihara, Takasuke Fukuhara, Chikako Ono, Satomi Yamamoto, Kentaro Uemura, Toru Okamoto, Masaya Sugiyama, Daisuke Motooka, Shota Nakamura, Masato Ikawa, Masashi Mizokami, Yoshihiko Maehara, Yoshiharu Matsuura

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number6122
JournalScientific reports
Volume7
Issue number1
DOIs
Publication statusPublished - Dec 1 2017

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Deoxyribonuclease I
Virus Replication
Hepatitis B virus
Genome
Guide RNA
DNA
Liver
Chronic Hepatitis B
Viral Proteins
Mutagenesis
Mutation

All Science Journal Classification (ASJC) codes

  • General

Cite this

Kurihara, T., Fukuhara, T., Ono, C., Yamamoto, S., Uemura, K., Okamoto, T., ... Matsuura, Y. (2017). Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9. Scientific reports, 7(1), [6122]. https://doi.org/10.1038/s41598-017-05905-w

Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9. / Kurihara, Takeshi; Fukuhara, Takasuke; Ono, Chikako; Yamamoto, Satomi; Uemura, Kentaro; Okamoto, Toru; Sugiyama, Masaya; Motooka, Daisuke; Nakamura, Shota; Ikawa, Masato; Mizokami, Masashi; Maehara, Yoshihiko; Matsuura, Yoshiharu.

In: Scientific reports, Vol. 7, No. 1, 6122, 01.12.2017.

Research output: Contribution to journalArticle

Kurihara, T, Fukuhara, T, Ono, C, Yamamoto, S, Uemura, K, Okamoto, T, Sugiyama, M, Motooka, D, Nakamura, S, Ikawa, M, Mizokami, M, Maehara, Y & Matsuura, Y 2017, 'Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9', Scientific reports, vol. 7, no. 1, 6122. https://doi.org/10.1038/s41598-017-05905-w
Kurihara T, Fukuhara T, Ono C, Yamamoto S, Uemura K, Okamoto T et al. Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9. Scientific reports. 2017 Dec 1;7(1). 6122. https://doi.org/10.1038/s41598-017-05905-w
Kurihara, Takeshi ; Fukuhara, Takasuke ; Ono, Chikako ; Yamamoto, Satomi ; Uemura, Kentaro ; Okamoto, Toru ; Sugiyama, Masaya ; Motooka, Daisuke ; Nakamura, Shota ; Ikawa, Masato ; Mizokami, Masashi ; Maehara, Yoshihiko ; Matsuura, Yoshiharu. / Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9. In: Scientific reports. 2017 ; Vol. 7, No. 1.
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