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, Masahito; Mizokami, Masashi; Maehara, Yoshihiko; Matsuura, Yoshiharu

doi: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 et al.

Scientific Reports, 2017, 7(1), 6122

Number of Access:2,185（2025-08-15 09:42 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/78577

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Title	Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9
Creator	CreatorName Kurihara, Takeshi
	NRID 1000070598739 1000070598739 CreatorName Fukuhara, Takasuke
	NRID 1000030626437 1000030626437 researchmap permalink ChikakoOno ChikakoOno Scopus Author ID 54990934700 54990934700 CreatorName Ono, Chikako
	CreatorName Yamamoto, Satomi
	CreatorName Uemura, Kentaro
	NRID 1000080628595 1000080628595 researchmap permalink TOKAMOTO TOKAMOTO Scopus Author ID 10144905800 10144905800 CreatorName Okamoto, Toru
	NRID 1000020612427 1000020612427 CreatorName Sugiyama, Masaya
	NRID 1000010636830 1000010636830 researchmap permalink daisukem daisukem Scopus Author ID 35315737900 35315737900 CreatorName Motooka, Daisuke
	NRID 1000090432434 1000090432434 ORCID 0000-0003-2058-5942 0000-0003-2058-5942 researchmap permalink nkmrshota nkmrshota Scopus Author ID 35315778200 35315778200 CreatorName Nakamura, Shota
	NRID 1000020304066 1000020304066 ORCID 0000-0001-9859-6217 0000-0001-9859-6217 researchmap permalink read0054910 read0054910 Scopus Author ID 16689054500 16689054500 CreatorName Ikawa, Masahito
	NRID 1000040166038 1000040166038 CreatorName Mizokami, Masashi
	NRID 1000080165662 1000080165662 CreatorName Maehara, Yoshihiko
	NRID 1000050157252 1000050157252 ORCID 0000-0001-9091-8285 0000-0001-9091-8285 researchmap permalink matsuura_123 matsuura_123 Scopus Author ID 7202988996 7202988996 CreatorName Matsuura, Yoshiharu
Description	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
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.
Publisher	Nature Research
Source Title	Scientific Reports
Volume (Issue)	7(1)
Page	6122
Date of Issued	2017-07-21
Language	English
Handle URL	https://hdl.handle.net/11094/78577
eISSN	20452322
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1038/s41598-017-05905-w
Relation.isIdenticalTo	PMID 28733609
Access Rights	open access
CopyRight	© 2017 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License.
oaire:version	Version of Record
Category	学術雑誌論文 Journal Article

Resource Type	journal article
Local Type	Journal Article
DCTERMS.bibliographicCitation	Scientific Reports. 2017, 7(1), p. 6122
DC.title	Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9
DC.creator	Kurihara, Takeshi
	Fukuhara, Takasuke
	Ono, Chikako
	Yamamoto, Satomi
	Uemura, Kentaro
	Okamoto, Toru
	Sugiyama, Masaya
	Motooka, Daisuke
	Nakamura, Shota
	Ikawa, Masahito
	Mizokami, Masashi
	Maehara, Yoshihiko
	Matsuura, Yoshiharu
DC.publisher	Nature Research
DC.language' scheme='DCTERMS.RFC1766	English
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2017-07-21
DC.identifier	https://doi.org/10.1038/s41598-017-05905-w
DC.description	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
DCTERMS.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.
DC.rights	© 2017 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	78577
citation_title	Suppression of HBV replication by the expression of nickase-and nuclease dead-Cas9
citation_author	Kurihara, Takeshi
	Fukuhara, Takasuke
	Ono, Chikako
	Yamamoto, Satomi
	Uemura, Kentaro
	Okamoto, Toru
	Sugiyama, Masaya
	Motooka, Daisuke
	Nakamura, Shota
	Ikawa, Masahito
	Mizokami, Masashi
	Maehara, Yoshihiko
	Matsuura, Yoshiharu
citation_publisher	Nature Research
citation_language	English
citation_date	2017-07-21
citation_journal_title	Scientific Reports
citation_volume	7
citation_issue	1
citation_firstpage	6122
citation_doi	https://doi.org/10.1038/s41598-017-05905-w
citation_pmid	28733609
citation_public_url	https://hdl.handle.net/11094/78577

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

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