Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice
Identifieur interne : 000758 ( Ncbi/Curation ); précédent : 000757; suivant : 000759Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice
Auteurs : Prasanta K. Dash [États-Unis] ; Rafal Kaminski [États-Unis] ; Ramona Bella [États-Unis] ; Hang Su [États-Unis] ; Saumi Mathews [États-Unis] ; Taha M. Ahooyi [États-Unis] ; Chen Chen [États-Unis] ; Pietro Mancuso [États-Unis] ; Rahsan Sariyer [États-Unis] ; Pasquale Ferrante [États-Unis] ; Martina Donadoni [États-Unis] ; Jake A. Robinson [États-Unis] ; Brady Sillman [États-Unis] ; Zhiyi Lin [États-Unis] ; James R. Hilaire [États-Unis] ; Mary Banoub [États-Unis] ; Monalisha Elango [États-Unis] ; Nagsen Gautam [États-Unis] ; R. Lee Mosley [États-Unis] ; Larisa Y. Poluektova [États-Unis] ; Joellyn Mcmillan [États-Unis] ; Aditya N. Bade [États-Unis] ; Santhi Gorantla [États-Unis] ; Ilker K. Sariyer [États-Unis] ; Tricia H. Burdo [États-Unis] ; Won-Bin Young [États-Unis] ; Shohreh Amini [États-Unis] ; Jennifer Gordon [États-Unis] ; Jeffrey M. Jacobson [États-Unis] ; Benson Edagwa [États-Unis] ; Kamel Khalili [États-Unis] ; Howard E. Gendelman [États-Unis]Source :
- Nature Communications [ 2041-1723 ] ; 2019.
Abstract
Elimination of HIV-1 requires clearance and removal of integrated proviral DNA from infected cells and tissues. Here, sequential long-acting slow-effective release antiviral therapy (LASER ART) and CRISPR-Cas9 demonstrate viral clearance in latent infectious reservoirs in HIV-1 infected humanized mice. HIV-1 subgenomic DNA fragments, spanning the long terminal repeats and the Gag gene, are excised in vivo, resulting in elimination of integrated proviral DNA; virus is not detected in blood, lymphoid tissue, bone marrow and brain by nested and digital-droplet PCR as well as RNAscope tests. No CRISPR-Cas9 mediated off-target effects are detected. Adoptive transfer of human immunocytes from dual treated, virus-free animals to uninfected humanized mice fails to produce infectious progeny virus. In contrast, HIV-1 is readily detected following sole LASER ART or CRISPR-Cas9 treatment. These data provide proof-of-concept that permanent viral elimination is possible.
Url:
DOI: 10.1038/s41467-019-10366-y
PubMed: 31266936
PubMed Central: 6606613
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<author><name sortKey="Gautam, Nagsen" sort="Gautam, Nagsen" uniqKey="Gautam N" first="Nagsen" last="Gautam">Nagsen Gautam</name>
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<author><name sortKey="Mosley, R Lee" sort="Mosley, R Lee" uniqKey="Mosley R" first="R. Lee" last="Mosley">R. Lee Mosley</name>
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<author><name sortKey="Mcmillan, Joellyn" sort="Mcmillan, Joellyn" uniqKey="Mcmillan J" first="Joellyn" last="Mcmillan">Joellyn Mcmillan</name>
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<author><name sortKey="Bade, Aditya N" sort="Bade, Aditya N" uniqKey="Bade A" first="Aditya N." last="Bade">Aditya N. Bade</name>
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<author><name sortKey="Gorantla, Santhi" sort="Gorantla, Santhi" uniqKey="Gorantla S" first="Santhi" last="Gorantla">Santhi Gorantla</name>
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<author><name sortKey="Sariyer, Ilker K" sort="Sariyer, Ilker K" uniqKey="Sariyer I" first="Ilker K." last="Sariyer">Ilker K. Sariyer</name>
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<author><name sortKey="Burdo, Tricia H" sort="Burdo, Tricia H" uniqKey="Burdo T" first="Tricia H." last="Burdo">Tricia H. Burdo</name>
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<author><name sortKey="Young, Won Bin" sort="Young, Won Bin" uniqKey="Young W" first="Won-Bin" last="Young">Won-Bin Young</name>
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<author><name sortKey="Amini, Shohreh" sort="Amini, Shohreh" uniqKey="Amini S" first="Shohreh" last="Amini">Shohreh Amini</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice</title>
<author><name sortKey="Dash, Prasanta K" sort="Dash, Prasanta K" uniqKey="Dash P" first="Prasanta K." last="Dash">Prasanta K. Dash</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
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<placeName><region type="state">Nebraska</region>
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</author>
<author><name sortKey="Kaminski, Rafal" sort="Kaminski, Rafal" uniqKey="Kaminski R" first="Rafal" last="Kaminski">Rafal Kaminski</name>
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<institution>Department of Neuroscience,</institution>
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Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Pennsylvanie</region>
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<author><name sortKey="Bella, Ramona" sort="Bella, Ramona" uniqKey="Bella R" first="Ramona" last="Bella">Ramona Bella</name>
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Philadelphia, PA 19115 USA</nlm:aff>
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<placeName><region type="state">Pennsylvanie</region>
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<author><name sortKey="Su, Hang" sort="Su, Hang" uniqKey="Su H" first="Hang" last="Su">Hang Su</name>
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<institution-id institution-id-type="GRID">grid.266813.8</institution-id>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
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<author><name sortKey="Mathews, Saumi" sort="Mathews, Saumi" uniqKey="Mathews S" first="Saumi" last="Mathews">Saumi Mathews</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
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<author><name sortKey="Ahooyi, Taha M" sort="Ahooyi, Taha M" uniqKey="Ahooyi T" first="Taha M." last="Ahooyi">Taha M. Ahooyi</name>
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<author><name sortKey="Chen, Chen" sort="Chen, Chen" uniqKey="Chen C" first="Chen" last="Chen">Chen Chen</name>
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<author><name sortKey="Mancuso, Pietro" sort="Mancuso, Pietro" uniqKey="Mancuso P" first="Pietro" last="Mancuso">Pietro Mancuso</name>
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Philadelphia, PA 19115 USA</nlm:aff>
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<placeName><region type="state">Pennsylvanie</region>
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<author><name sortKey="Sariyer, Rahsan" sort="Sariyer, Rahsan" uniqKey="Sariyer R" first="Rahsan" last="Sariyer">Rahsan Sariyer</name>
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<author><name sortKey="Ferrante, Pasquale" sort="Ferrante, Pasquale" uniqKey="Ferrante P" first="Pasquale" last="Ferrante">Pasquale Ferrante</name>
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Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Pennsylvanie</region>
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<author><name sortKey="Donadoni, Martina" sort="Donadoni, Martina" uniqKey="Donadoni M" first="Martina" last="Donadoni">Martina Donadoni</name>
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<institution-id institution-id-type="GRID">grid.264727.2</institution-id>
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Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
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<author><name sortKey="Robinson, Jake A" sort="Robinson, Jake A" uniqKey="Robinson J" first="Jake A." last="Robinson">Jake A. Robinson</name>
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<institution-id institution-id-type="GRID">grid.264727.2</institution-id>
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<author><name sortKey="Sillman, Brady" sort="Sillman, Brady" uniqKey="Sillman B" first="Brady" last="Sillman">Brady Sillman</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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<author><name sortKey="Lin, Zhiyi" sort="Lin, Zhiyi" uniqKey="Lin Z" first="Zhiyi" last="Lin">Zhiyi Lin</name>
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<institution-id institution-id-type="GRID">grid.266813.8</institution-id>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
</placeName>
<wicri:cityArea>Omaha</wicri:cityArea>
</affiliation>
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<author><name sortKey="Hilaire, James R" sort="Hilaire, James R" uniqKey="Hilaire J" first="James R." last="Hilaire">James R. Hilaire</name>
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<institution-id institution-id-type="GRID">grid.266813.8</institution-id>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
</placeName>
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</affiliation>
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<author><name sortKey="Banoub, Mary" sort="Banoub, Mary" uniqKey="Banoub M" first="Mary" last="Banoub">Mary Banoub</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
</placeName>
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</author>
<author><name sortKey="Elango, Monalisha" sort="Elango, Monalisha" uniqKey="Elango M" first="Monalisha" last="Elango">Monalisha Elango</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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</placeName>
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</affiliation>
</author>
<author><name sortKey="Gautam, Nagsen" sort="Gautam, Nagsen" uniqKey="Gautam N" first="Nagsen" last="Gautam">Nagsen Gautam</name>
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<institution>Department of Pharmaceutical Sciences,</institution>
<institution>College of Pharmacy, University of Nebraska Medical Center,</institution>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
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<author><name sortKey="Mosley, R Lee" sort="Mosley, R Lee" uniqKey="Mosley R" first="R. Lee" last="Mosley">R. Lee Mosley</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
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</affiliation>
</author>
<author><name sortKey="Poluektova, Larisa Y" sort="Poluektova, Larisa Y" uniqKey="Poluektova L" first="Larisa Y." last="Poluektova">Larisa Y. Poluektova</name>
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<institution>University of Nebraska Medical Center,</institution>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
</placeName>
<wicri:cityArea>Omaha</wicri:cityArea>
</affiliation>
</author>
<author><name sortKey="Mcmillan, Joellyn" sort="Mcmillan, Joellyn" uniqKey="Mcmillan J" first="Joellyn" last="Mcmillan">Joellyn Mcmillan</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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<author><name sortKey="Bade, Aditya N" sort="Bade, Aditya N" uniqKey="Bade A" first="Aditya N." last="Bade">Aditya N. Bade</name>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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<author><name sortKey="Gorantla, Santhi" sort="Gorantla, Santhi" uniqKey="Gorantla S" first="Santhi" last="Gorantla">Santhi Gorantla</name>
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<institution-id institution-id-type="GRID">grid.266813.8</institution-id>
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<institution>University of Nebraska Medical Center,</institution>
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Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
</placeName>
<wicri:cityArea>Omaha</wicri:cityArea>
</affiliation>
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<author><name sortKey="Sariyer, Ilker K" sort="Sariyer, Ilker K" uniqKey="Sariyer I" first="Ilker K." last="Sariyer">Ilker K. Sariyer</name>
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Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
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</affiliation>
</author>
<author><name sortKey="Burdo, Tricia H" sort="Burdo, Tricia H" uniqKey="Burdo T" first="Tricia H." last="Burdo">Tricia H. Burdo</name>
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<institution-id institution-id-type="GRID">grid.264727.2</institution-id>
<institution>Department of Neuroscience,</institution>
<institution>Lewis Katz School of Medicine at Temple University,</institution>
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Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Pennsylvanie</region>
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<author><name sortKey="Young, Won Bin" sort="Young, Won Bin" uniqKey="Young W" first="Won-Bin" last="Young">Won-Bin Young</name>
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<institution>Department of Neuroscience,</institution>
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Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
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<author><name sortKey="Amini, Shohreh" sort="Amini, Shohreh" uniqKey="Amini S" first="Shohreh" last="Amini">Shohreh Amini</name>
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Philadelphia, PA 19115 USA</nlm:aff>
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Philadelphia, PA 19115 USA</nlm:aff>
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<institution>Department of Neuroscience,</institution>
<institution>Lewis Katz School of Medicine at Temple University,</institution>
</institution-wrap>
Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
<wicri:cityArea>Philadelphia</wicri:cityArea>
</affiliation>
</author>
<author><name sortKey="Edagwa, Benson" sort="Edagwa, Benson" uniqKey="Edagwa B" first="Benson" last="Edagwa">Benson Edagwa</name>
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<institution-id institution-id-type="GRID">grid.266813.8</institution-id>
<institution>Department of Pharmacology and Experimental Neuroscience,</institution>
<institution>University of Nebraska Medical Center,</institution>
</institution-wrap>
Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
</placeName>
<wicri:cityArea>Omaha</wicri:cityArea>
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</author>
<author><name sortKey="Khalili, Kamel" sort="Khalili, Kamel" uniqKey="Khalili K" first="Kamel" last="Khalili">Kamel Khalili</name>
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<institution>Department of Neuroscience,</institution>
<institution>Lewis Katz School of Medicine at Temple University,</institution>
</institution-wrap>
Philadelphia, PA 19115 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
<wicri:cityArea>Philadelphia</wicri:cityArea>
</affiliation>
</author>
<author><name sortKey="Gendelman, Howard E" sort="Gendelman, Howard E" uniqKey="Gendelman H" first="Howard E." last="Gendelman">Howard E. Gendelman</name>
<affiliation wicri:level="2"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0001 0666 4105</institution-id>
<institution-id institution-id-type="GRID">grid.266813.8</institution-id>
<institution>Department of Pharmacology and Experimental Neuroscience,</institution>
<institution>University of Nebraska Medical Center,</institution>
</institution-wrap>
Omaha, NE 68198-5880 USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<placeName><region type="state">Nebraska</region>
</placeName>
<wicri:cityArea>Omaha</wicri:cityArea>
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</analytic>
<series><title level="j">Nature Communications</title>
<idno type="eISSN">2041-1723</idno>
<imprint><date when="2019">2019</date>
</imprint>
</series>
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<front><div type="abstract" xml:lang="en"><p id="Par1">Elimination of HIV-1 requires clearance and removal of integrated proviral DNA from infected cells and tissues. Here, sequential long-acting slow-effective release antiviral therapy (LASER ART) and CRISPR-Cas9 demonstrate viral clearance in latent infectious reservoirs in HIV-1 infected humanized mice. HIV-1 subgenomic DNA fragments, spanning the long terminal repeats and the Gag gene, are excised in vivo, resulting in elimination of integrated proviral DNA; virus is not detected in blood, lymphoid tissue, bone marrow and brain by nested and digital-droplet PCR as well as RNAscope tests. No CRISPR-Cas9 mediated off-target effects are detected. Adoptive transfer of human immunocytes from dual treated, virus-free animals to uninfected humanized mice fails to produce infectious progeny virus. In contrast, HIV-1 is readily detected following sole LASER ART or CRISPR-Cas9 treatment. These data provide proof-of-concept that permanent viral elimination is possible.</p>
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