CRISPR-mediated activation of endogenous BST-2/tetherin expression inhibits wild-type HIV-1 production
Identifieur interne : 000156 ( Pmc/Curation ); précédent : 000155; suivant : 000157CRISPR-mediated activation of endogenous BST-2/tetherin expression inhibits wild-type HIV-1 production
Auteurs : Yanzhao Zhang [Japon] ; Seiya Ozono [Japon] ; Weitong Yao [Japon] ; Minoru Tobiume [Japon] ; Shoji Yamaoka [Japon] ; Satoshi Kishigami [Japon] ; Hideaki Fujita [Japon] ; Kenzo Tokunaga [Japon]Source :
- Scientific Reports [ 2045-2322 ] ; 2019.
Abstract
The CRISPR technology not only can knock out target genes by using the RNA-guided Cas9 nuclease but also can activate their expression when a nuclease-deficient Cas9 (dCas9) is employed. Using the latter function, we here show the effect of the CRISPR-mediated pinpoint activation of endogenous expression of BST-2 (also known as tetherin), a virus restriction factor with a broad antiviral spectrum. Single-guide RNA (sgRNA) sequences targeting the BST-2 promoter were selected by promoter assays. Potential sgRNAs and dCas9 fused to the VP64 transactivation domain, along with an accessory transcriptional activator complex, were introduced into cells by lentiviral transduction. Increased expression of
Url:
DOI: 10.1038/s41598-019-40003-z
PubMed: 30816279
PubMed Central: 6395588
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wicri:step="Curation">000156</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">CRISPR-mediated activation of endogenous BST-2/tetherin expression inhibits wild-type HIV-1 production</title><author><name sortKey="Zhang, Yanzhao" sort="Zhang, Yanzhao" uniqKey="Zhang Y" first="Yanzhao" last="Zhang">Yanzhao Zhang</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2220 1880</institution-id><institution-id institution-id-type="GRID">grid.410795.e</institution-id><institution>Department of Pathology,</institution><institution>National Institute of Infectious Diseases,</institution></institution-wrap>Tokyo, 162-8640 Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Tokyo</wicri:regionArea></affiliation></author><author><name sortKey="Ozono, Seiya" sort="Ozono, Seiya" uniqKey="Ozono S" first="Seiya" last="Ozono">Seiya Ozono</name><affiliation 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xml:lang="fr">Japon</country><wicri:regionArea>Tokyo</wicri:regionArea></affiliation></author><author><name sortKey="Tobiume, Minoru" sort="Tobiume, Minoru" uniqKey="Tobiume M" first="Minoru" last="Tobiume">Minoru Tobiume</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2220 1880</institution-id><institution-id institution-id-type="GRID">grid.410795.e</institution-id><institution>Department of Pathology,</institution><institution>National Institute of Infectious Diseases,</institution></institution-wrap>Tokyo, 162-8640 Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Tokyo</wicri:regionArea></affiliation></author><author><name sortKey="Yamaoka, Shoji" sort="Yamaoka, Shoji" uniqKey="Yamaoka S" first="Shoji" last="Yamaoka">Shoji Yamaoka</name><affiliation wicri:level="1"><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 1014 9130</institution-id><institution-id institution-id-type="GRID">grid.265073.5</institution-id><institution>Department of Molecular Virology,</institution><institution>Tokyo Medical and Dental University,</institution></institution-wrap>Tokyo, 113-8519 Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Tokyo</wicri:regionArea></affiliation></author><author><name sortKey="Kishigami, Satoshi" sort="Kishigami, Satoshi" uniqKey="Kishigami S" first="Satoshi" last="Kishigami">Satoshi Kishigami</name><affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0291 3581</institution-id><institution-id institution-id-type="GRID">grid.267500.6</institution-id><institution>Faculty of Life and Environmental Sciences,</institution><institution>University of Yamanashi,</institution></institution-wrap>Yamanashi, 400-8510 Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Yamanashi</wicri:regionArea></affiliation></author><author><name sortKey="Fujita, Hideaki" sort="Fujita, Hideaki" uniqKey="Fujita H" first="Hideaki" last="Fujita">Hideaki Fujita</name><affiliation wicri:level="1"><nlm:aff id="Aff4"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0647 5488</institution-id><institution-id institution-id-type="GRID">grid.411871.a</institution-id><institution>Faculty of Pharmaceutical Sciences,</institution><institution>Nagasaki International University,</institution></institution-wrap>Nagasaki, 859-3298 Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Nagasaki</wicri:regionArea></affiliation></author><author><name sortKey="Tokunaga, Kenzo" sort="Tokunaga, Kenzo" uniqKey="Tokunaga K" first="Kenzo" last="Tokunaga">Kenzo Tokunaga</name><affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2220 1880</institution-id><institution-id institution-id-type="GRID">grid.410795.e</institution-id><institution>Department of Pathology,</institution><institution>National Institute of Infectious Diseases,</institution></institution-wrap>Tokyo, 162-8640 Japan</nlm:aff><country xml:lang="fr">Japon</country><wicri:regionArea>Tokyo</wicri:regionArea></affiliation></author></analytic><series><title level="j">Scientific Reports</title><idno type="eISSN">2045-2322</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="Par1">The CRISPR technology not only can knock out target genes by using the RNA-guided Cas9 nuclease but also can activate their expression when a nuclease-deficient Cas9 (dCas9) is employed. Using the latter function, we here show the effect of the CRISPR-mediated pinpoint activation of endogenous expression of BST-2 (also known as tetherin), a virus restriction factor with a broad antiviral spectrum. Single-guide RNA (sgRNA) sequences targeting the BST-2 promoter were selected by promoter assays. Potential sgRNAs and dCas9 fused to the VP64 transactivation domain, along with an accessory transcriptional activator complex, were introduced into cells by lentiviral transduction. Increased expression of <italic>BST-2</italic> mRNA in transduced cells was confirmed by real-time RT-PCR. Cells in which BST-2 expression was highly enhanced showed the effective inhibition of HIV-1 production and replication even in the presence of the viral antagonist Vpu against BST-2. These findings confirm that the physiological stoichiometry between host restriction factors and viral antagonists may determine the outcome of the battle with viruses.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Tokunaga, K" uniqKey="Tokunaga K">K Tokunaga</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sakai, H" uniqKey="Sakai H">H Sakai</name></author><author><name sortKey="Tokunaga, K" uniqKey="Tokunaga K">K Tokunaga</name></author><author><name sortKey="Kawamura, M" uniqKey="Kawamura M">M Kawamura</name></author><author><name sortKey="Adachi, A" uniqKey="Adachi A">A Adachi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Kawamura, M" uniqKey="Kawamura M">M Kawamura</name></author><author><name sortKey="Sakai, H" uniqKey="Sakai H">H Sakai</name></author><author><name sortKey="Adachi, A" uniqKey="Adachi A">A 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Sci Rep</journal-id><journal-id journal-id-type="iso-abbrev">Sci Rep</journal-id><journal-title-group><journal-title>Scientific Reports</journal-title></journal-title-group><issn pub-type="epub">2045-2322</issn><publisher><publisher-name>Nature Publishing Group UK</publisher-name><publisher-loc>London</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">30816279</article-id><article-id pub-id-type="pmc">6395588</article-id><article-id pub-id-type="publisher-id">40003</article-id><article-id pub-id-type="doi">10.1038/s41598-019-40003-z</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>CRISPR-mediated activation of endogenous BST-2/tetherin expression inhibits wild-type HIV-1 production</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Zhang</surname><given-names>Yanzhao</given-names></name><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Ozono</surname><given-names>Seiya</given-names></name><xref ref-type="aff" rid="Aff1">1</xref><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Yao</surname><given-names>Weitong</given-names></name><xref ref-type="aff" rid="Aff1">1</xref><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Tobiume</surname><given-names>Minoru</given-names></name><xref ref-type="aff" rid="Aff1">1</xref></contrib><contrib contrib-type="author"><name><surname>Yamaoka</surname><given-names>Shoji</given-names></name><xref ref-type="aff" rid="Aff3">3</xref></contrib><contrib contrib-type="author"><name><surname>Kishigami</surname><given-names>Satoshi</given-names></name><xref ref-type="aff" rid="Aff2">2</xref></contrib><contrib contrib-type="author"><name><surname>Fujita</surname><given-names>Hideaki</given-names></name><xref ref-type="aff" rid="Aff4">4</xref></contrib><contrib contrib-type="author" corresp="yes"><name><surname>Tokunaga</surname><given-names>Kenzo</given-names></name><address><email>tokunaga@nih.go.jp</email></address><xref ref-type="aff" rid="Aff1">1</xref></contrib><aff id="Aff1"><label>1</label><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 2220 1880</institution-id><institution-id institution-id-type="GRID">grid.410795.e</institution-id><institution>Department of Pathology,</institution><institution>National Institute of Infectious Diseases,</institution></institution-wrap>Tokyo, 162-8640 Japan</aff><aff id="Aff2"><label>2</label><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0291 3581</institution-id><institution-id institution-id-type="GRID">grid.267500.6</institution-id><institution>Faculty of Life and Environmental Sciences,</institution><institution>University of Yamanashi,</institution></institution-wrap>Yamanashi, 400-8510 Japan</aff><aff id="Aff3"><label>3</label><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 1014 9130</institution-id><institution-id institution-id-type="GRID">grid.265073.5</institution-id><institution>Department of Molecular Virology,</institution><institution>Tokyo Medical and Dental University,</institution></institution-wrap>Tokyo, 113-8519 Japan</aff><aff id="Aff4"><label>4</label><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0647 5488</institution-id><institution-id institution-id-type="GRID">grid.411871.a</institution-id><institution>Faculty of Pharmaceutical Sciences,</institution><institution>Nagasaki International University,</institution></institution-wrap>Nagasaki, 859-3298 Japan</aff></contrib-group><pub-date pub-type="epub"><day>28</day><month>2</month><year>2019</year></pub-date><pub-date pub-type="pmc-release"><day>28</day><month>2</month><year>2019</year></pub-date><pub-date pub-type="collection"><year>2019</year></pub-date><volume>9</volume><elocation-id>3134</elocation-id><history><date date-type="received"><day>14</day><month>11</month><year>2018</year></date><date date-type="accepted"><day>7</day><month>2</month><year>2019</year></date></history><permissions><copyright-statement>© The Author(s) 2019</copyright-statement><license license-type="OpenAccess"><license-p><bold>Open Access</bold> 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 <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>.</license-p></license></permissions><abstract id="Abs1"><p id="Par1">The CRISPR technology not only can knock out target genes by using the RNA-guided Cas9 nuclease but also can activate their expression when a nuclease-deficient Cas9 (dCas9) is employed. Using the latter function, we here show the effect of the CRISPR-mediated pinpoint activation of endogenous expression of BST-2 (also known as tetherin), a virus restriction factor with a broad antiviral spectrum. Single-guide RNA (sgRNA) sequences targeting the BST-2 promoter were selected by promoter assays. Potential sgRNAs and dCas9 fused to the VP64 transactivation domain, along with an accessory transcriptional activator complex, were introduced into cells by lentiviral transduction. Increased expression of <italic>BST-2</italic> mRNA in transduced cells was confirmed by real-time RT-PCR. Cells in which BST-2 expression was highly enhanced showed the effective inhibition of HIV-1 production and replication even in the presence of the viral antagonist Vpu against BST-2. These findings confirm that the physiological stoichiometry between host restriction factors and viral antagonists may determine the outcome of the battle with viruses.</p></abstract><funding-group><award-group><funding-source><institution-wrap><institution-id institution-id-type="FundRef">https://doi.org/10.13039/100009619</institution-id><institution>Japan Agency for Medical Research and Development (AMED)</institution></institution-wrap></funding-source><award-id>17fk0410308j0503</award-id><principal-award-recipient><name><surname>Tokunaga</surname><given-names>Kenzo</given-names></name></principal-award-recipient></award-group></funding-group><funding-group><award-group><funding-source><institution-wrap><institution-id institution-id-type="FundRef">https://doi.org/10.13039/501100001691</institution-id><institution>MEXT | Japan Society for the Promotion of Science (JSPS)</institution></institution-wrap></funding-source><award-id>18K07156</award-id><principal-award-recipient><name><surname>Tokunaga</surname><given-names>Kenzo</given-names></name></principal-award-recipient></award-group></funding-group><custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name><meta-value>© The Author(s) 2019</meta-value></custom-meta></custom-meta-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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