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- Publisher Website: 10.1038/9886
- Scopus: eid_2-s2.0-0033021533
- PMID: 10385323
- WOS: WOS:000080716500030
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Article: Integration complexes derived from HIV vectors for rapid assays in vitro
Title | Integration complexes derived from HIV vectors for rapid assays in vitro |
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Authors | |
Keywords | AIDS HIV Integrase Quantitative PCR Retroviral vector TaqMan |
Issue Date | 1999 |
Citation | Nature Biotechnology, 1999, v. 17, n. 6, p. 578-582 How to Cite? |
Abstract | Of three enzymes encoded by HIV-reverse transcriptase, protease, and integrase - only the first two have been exploited clinically as inhibitor targets. Efforts to develop inhibitors of purified integrase protein have yielded many compounds, but none with clinical utility. A different source of integration activity for studies in vitro is provided by replication intermediates isolated from HIV-infected cells. These preintegration complexes (PICs) can direct integration of the endogenously synthesized viral cDNA into an added target DNA in vitro. Despite their authentic activities, assays of PICs have not been widely used due to technical obstacles, particularly the requirement for handling large amounts of infectious HIV. Here, we describe greatly improved methods for producing PICs using HIV- based vectors that are capable of establishing an integrated provirus but not a spreading infection. We also report the development of a PIC integration assay using DNA-coated microtiter plates, which speeds assays of PIC integration in vitro. We used this method to screen a library of chemicals related to known integrase inhibitors and found a new compound, quinalizarin sulfate, that displayed enhanced activity against PICs. |
Persistent Identifier | http://hdl.handle.net/10722/341042 |
ISSN | 2023 Impact Factor: 33.1 2023 SCImago Journal Rankings: 18.117 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Hansen, Mark S.T. | - |
dc.contributor.author | Smith, George J. | - |
dc.contributor.author | Kafri, Tal | - |
dc.contributor.author | Molteni, Valentina | - |
dc.contributor.author | Siegel, Jay S. | - |
dc.contributor.author | Bushman, Frederic D. | - |
dc.date.accessioned | 2024-03-13T08:39:42Z | - |
dc.date.available | 2024-03-13T08:39:42Z | - |
dc.date.issued | 1999 | - |
dc.identifier.citation | Nature Biotechnology, 1999, v. 17, n. 6, p. 578-582 | - |
dc.identifier.issn | 1087-0156 | - |
dc.identifier.uri | http://hdl.handle.net/10722/341042 | - |
dc.description.abstract | Of three enzymes encoded by HIV-reverse transcriptase, protease, and integrase - only the first two have been exploited clinically as inhibitor targets. Efforts to develop inhibitors of purified integrase protein have yielded many compounds, but none with clinical utility. A different source of integration activity for studies in vitro is provided by replication intermediates isolated from HIV-infected cells. These preintegration complexes (PICs) can direct integration of the endogenously synthesized viral cDNA into an added target DNA in vitro. Despite their authentic activities, assays of PICs have not been widely used due to technical obstacles, particularly the requirement for handling large amounts of infectious HIV. Here, we describe greatly improved methods for producing PICs using HIV- based vectors that are capable of establishing an integrated provirus but not a spreading infection. We also report the development of a PIC integration assay using DNA-coated microtiter plates, which speeds assays of PIC integration in vitro. We used this method to screen a library of chemicals related to known integrase inhibitors and found a new compound, quinalizarin sulfate, that displayed enhanced activity against PICs. | - |
dc.language | eng | - |
dc.relation.ispartof | Nature Biotechnology | - |
dc.subject | AIDS | - |
dc.subject | HIV | - |
dc.subject | Integrase | - |
dc.subject | Quantitative PCR | - |
dc.subject | Retroviral vector | - |
dc.subject | TaqMan | - |
dc.title | Integration complexes derived from HIV vectors for rapid assays in vitro | - |
dc.type | Article | - |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1038/9886 | - |
dc.identifier.pmid | 10385323 | - |
dc.identifier.scopus | eid_2-s2.0-0033021533 | - |
dc.identifier.volume | 17 | - |
dc.identifier.issue | 6 | - |
dc.identifier.spage | 578 | - |
dc.identifier.epage | 582 | - |
dc.identifier.isi | WOS:000080716500030 | - |