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Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool

Effective gene therapy depends on the efficient transfer of therapeutic genes to target cells. None of the current technologies, however, satisfy all of the requirements necessary for gene therapy, because the plasma and nuclear membranes of mammalian cells are tight barriers against gene transfer u...

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Autores principales: Hashida, H, Miyamoto, M, Cho, Y, Hida, Y, Kato, K, Kurokawa, T, Okushiba, S, Kondo, S, Dosaka-Akita, H, Katoh, H
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 2004
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2409656/
https://www.ncbi.nlm.nih.gov/pubmed/15026809
http://dx.doi.org/10.1038/sj.bjc.6601680
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author Hashida, H
Miyamoto, M
Cho, Y
Hida, Y
Kato, K
Kurokawa, T
Okushiba, S
Kondo, S
Dosaka-Akita, H
Katoh, H
author_facet Hashida, H
Miyamoto, M
Cho, Y
Hida, Y
Kato, K
Kurokawa, T
Okushiba, S
Kondo, S
Dosaka-Akita, H
Katoh, H
author_sort Hashida, H
collection PubMed
description Effective gene therapy depends on the efficient transfer of therapeutic genes to target cells. None of the current technologies, however, satisfy all of the requirements necessary for gene therapy, because the plasma and nuclear membranes of mammalian cells are tight barriers against gene transfer using synthetic delivery systems. The protein transduction domain (PTD) of human immunodeficiency virus type 1 (HIV-1) Tat protein greatly facilitates protein transfer via membrane destabilisation. We synthesised polylysine peptides containing Tat PTD (TAT-pK), or other sequences, and investigated their potential as agents for gene transfer. The synthesised polypeptide TAT-pK retains DNA binding function and mediates delivery of a reporter gene to cultured cells. RGD motif binds with low affinity to alpha integrins which induce cell activation. Two control polypeptides, GGG-pK and RGD-pK, were synthesised and tested, but their gene transfer abilities were weaker than those of TAT-pK. TAT-pK-mediated gene transfer was enhanced in the presence of chloroquine or ammonium chloride, to a greater extent than that of cationic lipid-mediated gene transfer in most cancer cell lines tested. These data suggest that TAT-pK may be a potent candidate delivery vehicle that promotes gene transfer, dependent on the endocytic pathway. We conclude that the TAT-pK/DNA complex is useful as a minimal unit to package therapeutic genes and to transduce them into mammalian cells.
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spelling pubmed-24096562009-09-10 Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool Hashida, H Miyamoto, M Cho, Y Hida, Y Kato, K Kurokawa, T Okushiba, S Kondo, S Dosaka-Akita, H Katoh, H Br J Cancer Experimental Therapeutics Effective gene therapy depends on the efficient transfer of therapeutic genes to target cells. None of the current technologies, however, satisfy all of the requirements necessary for gene therapy, because the plasma and nuclear membranes of mammalian cells are tight barriers against gene transfer using synthetic delivery systems. The protein transduction domain (PTD) of human immunodeficiency virus type 1 (HIV-1) Tat protein greatly facilitates protein transfer via membrane destabilisation. We synthesised polylysine peptides containing Tat PTD (TAT-pK), or other sequences, and investigated their potential as agents for gene transfer. The synthesised polypeptide TAT-pK retains DNA binding function and mediates delivery of a reporter gene to cultured cells. RGD motif binds with low affinity to alpha integrins which induce cell activation. Two control polypeptides, GGG-pK and RGD-pK, were synthesised and tested, but their gene transfer abilities were weaker than those of TAT-pK. TAT-pK-mediated gene transfer was enhanced in the presence of chloroquine or ammonium chloride, to a greater extent than that of cationic lipid-mediated gene transfer in most cancer cell lines tested. These data suggest that TAT-pK may be a potent candidate delivery vehicle that promotes gene transfer, dependent on the endocytic pathway. We conclude that the TAT-pK/DNA complex is useful as a minimal unit to package therapeutic genes and to transduce them into mammalian cells. Nature Publishing Group 2004-03-22 2004-02-24 /pmc/articles/PMC2409656/ /pubmed/15026809 http://dx.doi.org/10.1038/sj.bjc.6601680 Text en Copyright © 2004 Cancer Research UK https://creativecommons.org/licenses/by/4.0/This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material.If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/.
spellingShingle Experimental Therapeutics
Hashida, H
Miyamoto, M
Cho, Y
Hida, Y
Kato, K
Kurokawa, T
Okushiba, S
Kondo, S
Dosaka-Akita, H
Katoh, H
Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool
title Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool
title_full Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool
title_fullStr Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool
title_full_unstemmed Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool
title_short Fusion of HIV-1 Tat protein transduction domain to poly-lysine as a new DNA delivery tool
title_sort fusion of hiv-1 tat protein transduction domain to poly-lysine as a new dna delivery tool
topic Experimental Therapeutics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2409656/
https://www.ncbi.nlm.nih.gov/pubmed/15026809
http://dx.doi.org/10.1038/sj.bjc.6601680
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