CRISPR-mediated activation of endogenous BST-2/tetherin expression inhibits wild-type HIV-1 production
Identifieur interne : 000669 ( Main/Exploration ); précédent : 000668; suivant : 000670CRISPR-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
Affiliations:
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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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Goffinet</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Shindo, K" uniqKey="Shindo K">K Shindo</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Mariani, R" uniqKey="Mariani R">R Mariani</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Sandler, Ng" uniqKey="Sandler N">NG Sandler</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Stoddart, Ca" uniqKey="Stoddart C">CA Stoddart</name></author><author><name sortKey="Keir, Me" uniqKey="Keir M">ME Keir</name></author><author><name sortKey="Mccune, Jm" uniqKey="Mccune J">JM McCune</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Fernandez, S" uniqKey="Fernandez S">S Fernandez</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Haas, Dw" uniqKey="Haas D">DW Haas</name></author></analytic></biblStruct><biblStruct><analytic><author><name 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Liao</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Labun, K" uniqKey="Labun K">K Labun</name></author><author><name sortKey="Montague, Tg" uniqKey="Montague T">TG Montague</name></author><author><name sortKey="Gagnon, Ja" uniqKey="Gagnon J">JA Gagnon</name></author><author><name sortKey="Thyme, Sb" uniqKey="Thyme S">SB Thyme</name></author><author><name sortKey="Valen, E" uniqKey="Valen E">E Valen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Bego, Mg" uniqKey="Bego M">MG Bego</name></author><author><name sortKey="Mercier, J" uniqKey="Mercier J">J Mercier</name></author><author><name sortKey="Cohen, Ea" uniqKey="Cohen E">ÉA Cohen</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dominguez, Aa" uniqKey="Dominguez A">AA Dominguez</name></author><author><name sortKey="Lim, Wa" uniqKey="Lim W">WA Lim</name></author><author><name sortKey="Qi, Ls" uniqKey="Qi L">LS Qi</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Gilbert, La" uniqKey="Gilbert L">LA Gilbert</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Iwabu, Y" uniqKey="Iwabu Y">Y Iwabu</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tada, T" uniqKey="Tada T">T Tada</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Masuyama, N" uniqKey="Masuyama N">N Masuyama</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>Japon</li></country></list><tree><country name="Japon"><noRegion><name sortKey="Zhang, Yanzhao" sort="Zhang, Yanzhao" uniqKey="Zhang Y" first="Yanzhao" last="Zhang">Yanzhao Zhang</name></noRegion><name sortKey="Fujita, Hideaki" sort="Fujita, Hideaki" uniqKey="Fujita H" first="Hideaki" last="Fujita">Hideaki Fujita</name><name sortKey="Kishigami, Satoshi" sort="Kishigami, Satoshi" uniqKey="Kishigami S" first="Satoshi" last="Kishigami">Satoshi Kishigami</name><name sortKey="Ozono, Seiya" sort="Ozono, Seiya" uniqKey="Ozono S" first="Seiya" last="Ozono">Seiya Ozono</name><name sortKey="Ozono, Seiya" sort="Ozono, Seiya" uniqKey="Ozono S" first="Seiya" last="Ozono">Seiya Ozono</name><name sortKey="Tobiume, Minoru" sort="Tobiume, Minoru" uniqKey="Tobiume M" first="Minoru" last="Tobiume">Minoru Tobiume</name><name sortKey="Tokunaga, Kenzo" sort="Tokunaga, Kenzo" uniqKey="Tokunaga K" first="Kenzo" last="Tokunaga">Kenzo Tokunaga</name><name sortKey="Yamaoka, Shoji" sort="Yamaoka, Shoji" uniqKey="Yamaoka S" first="Shoji" last="Yamaoka">Shoji Yamaoka</name><name sortKey="Yao, Weitong" sort="Yao, Weitong" uniqKey="Yao W" first="Weitong" last="Yao">Weitong Yao</name><name sortKey="Yao, Weitong" sort="Yao, Weitong" uniqKey="Yao W" first="Weitong" last="Yao">Weitong Yao</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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