Quantification of unintegrated HIV-1 DNA at the single cell level in vivo

In the nucleus of HIV-1 infected cells, unintegrated HIV-1 DNA molecules exist in the form of one and two LTR circles and linear molecules with degraded extremities. In tissue culture they are invariably more numerous than the provirus, the relative proportion of integrated to unintegrated forms var...

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Detalhes bibliográficos
Autores: Meyerhans, Andreas, Suspène, Rodolphe
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2012
País:España
Recursos:Universitat Pompeu Fabra
Repositório:Repositorio Digital de la UPF
OAI Identifier:oai:repositori.upf.edu:10230/23709
Acesso em linha:http://hdl.handle.net/10230/23709
http://dx.doi.org/10.1371/journal.pone.0036246
Access Level:Acceso aberto
Palavra-chave:Cèl·lules
VIH (Virus)
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spelling Quantification of unintegrated HIV-1 DNA at the single cell level in vivoMeyerhans, AndreasSuspène, RodolpheCèl·lulesVIH (Virus)ADNIn the nucleus of HIV-1 infected cells, unintegrated HIV-1 DNA molecules exist in the form of one and two LTR circles and linear molecules with degraded extremities. In tissue culture they are invariably more numerous than the provirus, the relative proportion of integrated to unintegrated forms varies widely from 1:1 to 1:10 and even over 1:100. In vivo, this ratio is unknown. To determine it, single nuclei from two infected patients with a known provirus copy number were microdissected, HIV DNA was amplified by nested PCR, cloned and individual clones sequenced. Given the extraordinary sequence complexity, we made the assumption that the total number of distinct sequences approximated to real number of amplifiable HIV-1 DNA templates in the nucleus. We found that the number of unintegrated DNA molecules increased linearly with the proviral copy number there being on average 86 unintegrated molecules per provirus.RS was supported by an EMBO Long Term Fellowship. AM was supported by grants from the Deutsche Forschungsgemeinschaft and the Spanish Ministry of Science and Innovation (SAF2010-21336)Public Library of Science (PLoS)201520152012info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfapplication/pdfhttp://hdl.handle.net/10230/23709http://dx.doi.org/10.1371/journal.pone.0036246reponame:Repositorio Digital de la UPFinstname:Universitat Pompeu FabraInglésPLoS One. 2012; 7(5): e36246info:eu-repo/grantAgreement/ES/3PN/SAF2010-21336© 2012 Suspène, Meyerhans. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedhttps://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repositori.upf.edu:10230/237092026-06-12T07:21:37Z
dc.title.none.fl_str_mv Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
title Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
spellingShingle Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
Meyerhans, Andreas
Cèl·lules
VIH (Virus)
ADN
title_short Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
title_full Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
title_fullStr Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
title_full_unstemmed Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
title_sort Quantification of unintegrated HIV-1 DNA at the single cell level in vivo
dc.creator.none.fl_str_mv Meyerhans, Andreas
Suspène, Rodolphe
author Meyerhans, Andreas
author_facet Meyerhans, Andreas
Suspène, Rodolphe
author_role author
author2 Suspène, Rodolphe
author2_role author
dc.subject.none.fl_str_mv Cèl·lules
VIH (Virus)
ADN
topic Cèl·lules
VIH (Virus)
ADN
description In the nucleus of HIV-1 infected cells, unintegrated HIV-1 DNA molecules exist in the form of one and two LTR circles and linear molecules with degraded extremities. In tissue culture they are invariably more numerous than the provirus, the relative proportion of integrated to unintegrated forms varies widely from 1:1 to 1:10 and even over 1:100. In vivo, this ratio is unknown. To determine it, single nuclei from two infected patients with a known provirus copy number were microdissected, HIV DNA was amplified by nested PCR, cloned and individual clones sequenced. Given the extraordinary sequence complexity, we made the assumption that the total number of distinct sequences approximated to real number of amplifiable HIV-1 DNA templates in the nucleus. We found that the number of unintegrated DNA molecules increased linearly with the proviral copy number there being on average 86 unintegrated molecules per provirus.
publishDate 2012
dc.date.none.fl_str_mv 2012
2015
2015
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10230/23709
http://dx.doi.org/10.1371/journal.pone.0036246
url http://hdl.handle.net/10230/23709
http://dx.doi.org/10.1371/journal.pone.0036246
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv PLoS One. 2012; 7(5): e36246
info:eu-repo/grantAgreement/ES/3PN/SAF2010-21336
dc.rights.none.fl_str_mv https://creativecommons.org/licenses/by/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science (PLoS)
publisher.none.fl_str_mv Public Library of Science (PLoS)
dc.source.none.fl_str_mv reponame:Repositorio Digital de la UPF
instname:Universitat Pompeu Fabra
instname_str Universitat Pompeu Fabra
reponame_str Repositorio Digital de la UPF
collection Repositorio Digital de la UPF
repository.name.fl_str_mv
repository.mail.fl_str_mv
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