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:	Lakshmikanth Mariyanna, Poornima Priyadarshini, Helga Hofmann-Sieber, Marcel Krepstakies, Nicole Walz, Adam Grundhoff, Frank Buchholz, Eberhard Hildt, Joachim Hauber
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/443823fc35f144baa7e568151a2cd991
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spelling	oai:doaj.org-article:443823fc35f144baa7e568151a2cd9912021-11-18T07:28:25ZExcision of HIV-1 proviral DNA by recombinant cell permeable tre-recombinase.1932-620310.1371/journal.pone.0031576https://doaj.org/article/443823fc35f144baa7e568151a2cd9912012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22348110/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Over 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.Lakshmikanth MariyannaPoornima PriyadarshiniHelga Hofmann-SieberMarcel KrepstakiesNicole WalzAdam GrundhoffFrank BuchholzEberhard HildtJoachim HauberPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 2, p e31576 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Lakshmikanth Mariyanna Poornima Priyadarshini Helga Hofmann-Sieber Marcel Krepstakies Nicole Walz Adam Grundhoff Frank Buchholz Eberhard Hildt Joachim Hauber Excision of HIV-1 proviral DNA by recombinant cell permeable tre-recombinase.
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.
format	article
author	Lakshmikanth Mariyanna Poornima Priyadarshini Helga Hofmann-Sieber Marcel Krepstakies Nicole Walz Adam Grundhoff Frank Buchholz Eberhard Hildt Joachim Hauber
author_facet	Lakshmikanth Mariyanna Poornima Priyadarshini Helga Hofmann-Sieber Marcel Krepstakies Nicole Walz Adam Grundhoff Frank Buchholz Eberhard Hildt Joachim Hauber
author_sort	Lakshmikanth Mariyanna
title	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_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_sort	excision of hiv-1 proviral dna by recombinant cell permeable tre-recombinase.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/443823fc35f144baa7e568151a2cd991
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Excision of HIV-1 proviral DNA by recombinant cell permeable tre-recombinase.

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