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https://hdl.handle.net/20.500.14094/90004907
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2024-07-27
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90004907 (fulltext)
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90004907
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open access
出版タイプ
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タイトル
CRISPR/Cas9 system targeting regulatory genes of HIV-1 inhibits viral replication in infected T-cell cultures
著者
Ophinni, Youdiil ; Inoue, Mari ; Kotaki, Tomohiro ; Kameoka, Masanori
著者名
Ophinni, Youdiil
著者名
Inoue, Mari
著者ID
A1352
研究者ID
1000010758816
KUID
https://kuid-rm-web.ofc.kobe-u.ac.jp/search/detail?systemId=849783d0576b0945520e17560c007669
著者名
Kotaki, Tomohiro
小瀧, 将裕
コタキ, トモヒロ
所属機関名
保健学研究科
著者ID
A0045
研究者ID
1000060281838
KUID
https://kuid-rm-web.ofc.kobe-u.ac.jp/search/detail?systemId=01cae9e1f2290a00520e17560c007669
著者名
Kameoka, Masanori
亀岡, 正典
カメオカ, マサノリ
所属機関名
保健学研究科
言語
English (英語)
収録物名
Scientific Reports
巻(号)
8
ページ
7784-7784
出版者
Nature Publishing Group
刊行日
2018-05-17
公開日
2018-05-30
抄録
The CRISPR/Cas9 system provides a novel and promising tool for editing the HIV-1 proviral genome. We designed RNA-guided CRISPR/Cas9 targeting the HIV-1 regulatory genes tat and rev with guide RNAs (gRNA) selected from each gene based on CRISPR specificity and sequence conservation across six major HIV-1 subtypes. Each gRNA was cloned into lentiCRISPRv2 before co-transfection to create a lentiviral vector and transduction into target cells. CRISPR/Cas9 transduction into 293T and HeLa cells stably expressing Tat and Rev proteins successfully abolished the expression of each protein relative to that in non-transduced and gRNA-absent vector-transduced cells. Tat functional assays showed significantly reduced HIV-1 promoter-driven luciferase expression after tat-CRISPR transduction, while Rev functional assays revealed abolished gpl20 expression after rev-CRISPR transduction. The target gene was mutated at the Cas9 cleavage site with high frequency and various indel mutations. Conversely, no mutations were detected at off-target sites and Cas9 expression had no effect on cell viability. CRISPR/Cas9 was further tested in persistently and latently HIV-l-infected T-cell lines, in which p24 levels were significantly suppressed even after cytokine reactivation, and multiplexing all six gRNAs further increased efficiency. Thus, the CRISPR/Cas9 system targeting HIV-1 regulatory genes may serve as a favorable means to achieve functional cures.
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保健学研究科
学術雑誌論文
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© 2018 Moriguchi et al.
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
関連情報
DOI
https://doi.org/10.1038/s41598-018-26190-1
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資源タイプ
journal article
eISSN
2045-2322
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