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Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase

Over the previous years, comprehensive studies on antiretroviral drugs resulted in the successful introduction of highly active antiretroviral therapy (HAART) into clinical practice for treatment of HIV/AIDS. However, there is still need for new therapeutic approaches, since HAART cannot eradicate H...

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Autores principales: Mariyanna, Lakshmikanth, Priyadarshini, Poornima, Hofmann-Sieber, Helga, Krepstakies, Marcel, Walz, Nicole, Grundhoff, Adam, Buchholz, Frank, Hildt, Eberhard, Hauber, Joachim
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3278460/
https://www.ncbi.nlm.nih.gov/pubmed/22348110
http://dx.doi.org/10.1371/journal.pone.0031576
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author Mariyanna, Lakshmikanth
Priyadarshini, Poornima
Hofmann-Sieber, Helga
Krepstakies, Marcel
Walz, Nicole
Grundhoff, Adam
Buchholz, Frank
Hildt, Eberhard
Hauber, Joachim
author_facet Mariyanna, Lakshmikanth
Priyadarshini, Poornima
Hofmann-Sieber, Helga
Krepstakies, Marcel
Walz, Nicole
Grundhoff, Adam
Buchholz, Frank
Hildt, Eberhard
Hauber, Joachim
author_sort Mariyanna, Lakshmikanth
collection PubMed
description Over the previous years, comprehensive studies on antiretroviral drugs resulted in the successful introduction of highly active antiretroviral therapy (HAART) into clinical practice for treatment of HIV/AIDS. However, there is still need for new therapeutic approaches, since HAART cannot eradicate HIV-1 from the infected organism and, unfortunately, can be associated with long-term toxicity and the development of drug resistance. In contrast, novel gene therapy strategies may have the potential to reverse the infection by eradicating HIV-1. For example, expression of long terminal repeat (LTR)-specific recombinase (Tre-recombinase) has been shown to result in chromosomal excision of proviral DNA and, in consequence, in the eradication of HIV-1 from infected cell cultures. However, the delivery of Tre-recombinase currently depends on the genetic manipulation of target cells, a process that is complicating such therapeutic approaches and, thus, might be undesirable in a clinical setting. In this report we demonstrate that E.coli expressed Tre-recombinases, tagged either with the protein transduction domain (PTD) from the HIV-1 Tat trans-activator or the translocation motif (TLM) of the Hepatitis B virus PreS2 protein, were able to translocate efficiently into cells and showed significant recombination activity on HIV-1 LTR sequences. Tre activity was observed using episomal and stable integrated reporter constructs in transfected HeLa cells. Furthermore, the TLM-tagged enzyme was able to excise the full-length proviral DNA from chromosomal integration sites of HIV-1-infected HeLa and CEM-SS cells. The presented data confirm Tre-recombinase activity on integrated HIV-1 and provide the basis for the non-genetic transient application of engineered recombinases, which may be a valuable component of future HIV eradication strategies.
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spelling pubmed-32784602012-02-17 Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase Mariyanna, Lakshmikanth Priyadarshini, Poornima Hofmann-Sieber, Helga Krepstakies, Marcel Walz, Nicole Grundhoff, Adam Buchholz, Frank Hildt, Eberhard Hauber, Joachim PLoS One Research Article Over the previous years, comprehensive studies on antiretroviral drugs resulted in the successful introduction of highly active antiretroviral therapy (HAART) into clinical practice for treatment of HIV/AIDS. However, there is still need for new therapeutic approaches, since HAART cannot eradicate HIV-1 from the infected organism and, unfortunately, can be associated with long-term toxicity and the development of drug resistance. In contrast, novel gene therapy strategies may have the potential to reverse the infection by eradicating HIV-1. For example, expression of long terminal repeat (LTR)-specific recombinase (Tre-recombinase) has been shown to result in chromosomal excision of proviral DNA and, in consequence, in the eradication of HIV-1 from infected cell cultures. However, the delivery of Tre-recombinase currently depends on the genetic manipulation of target cells, a process that is complicating such therapeutic approaches and, thus, might be undesirable in a clinical setting. In this report we demonstrate that E.coli expressed Tre-recombinases, tagged either with the protein transduction domain (PTD) from the HIV-1 Tat trans-activator or the translocation motif (TLM) of the Hepatitis B virus PreS2 protein, were able to translocate efficiently into cells and showed significant recombination activity on HIV-1 LTR sequences. Tre activity was observed using episomal and stable integrated reporter constructs in transfected HeLa cells. Furthermore, the TLM-tagged enzyme was able to excise the full-length proviral DNA from chromosomal integration sites of HIV-1-infected HeLa and CEM-SS cells. The presented data confirm Tre-recombinase activity on integrated HIV-1 and provide the basis for the non-genetic transient application of engineered recombinases, which may be a valuable component of future HIV eradication strategies. Public Library of Science 2012-02-13 /pmc/articles/PMC3278460/ /pubmed/22348110 http://dx.doi.org/10.1371/journal.pone.0031576 Text en Mariyanna et al. http://creativecommons.org/licenses/by/4.0/ 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 properly credited.
spellingShingle Research Article
Mariyanna, Lakshmikanth
Priyadarshini, Poornima
Hofmann-Sieber, Helga
Krepstakies, Marcel
Walz, Nicole
Grundhoff, Adam
Buchholz, Frank
Hildt, Eberhard
Hauber, Joachim
Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase
title Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase
title_full Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase
title_fullStr Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase
title_full_unstemmed Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase
title_short Excision of HIV-1 Proviral DNA by Recombinant Cell Permeable Tre-Recombinase
title_sort excision of hiv-1 proviral dna by recombinant cell permeable tre-recombinase
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3278460/
https://www.ncbi.nlm.nih.gov/pubmed/22348110
http://dx.doi.org/10.1371/journal.pone.0031576
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