Suppression of HBV replication by the expression of nickase- and nuclease dead-Cas9
| Autor: | Yoshiharu Matsuura, Daisuke Motooka, Takasuke Fukuhara, Kentaro Uemura, Satomi Yamamoto, Masaya Sugiyama, Shota Nakamura, Yoshihiko Maehara, Toru Okamoto, Takeshi Kurihara, Masashi Mizokami, Chikako Ono, Masato Ikawa |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Gene Expression Regulation Viral HBV RNA encapsidation signal epsilon Hepatitis B virus Science Gene Expression Genome Viral Biology medicine.disease_cause Virus Replication Genome Article 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine CRISPR-Associated Protein 9 medicine Deoxyribonuclease I Humans Subgenomic mRNA Nuclease Multidisciplinary Cas9 virus diseases Reproducibility of Results Virology Molecular biology digestive system diseases 030104 developmental biology Viral replication chemistry 030220 oncology & carcinogenesis DNA Viral Gene Targeting biology.protein Medicine DNA RNA Guide Kinetoplastida |
| Zdroj: | Scientific Reports Scientific Reports, Vol 7, Iss 1, Pp 1-13 (2017) |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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