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Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection

[Image: see text] The recent advances in genetic engineering demand the development of conceptually new methods to prepare and identify efficient vectors for the intracellular delivery of different nucleotide payloads ranging from short single-stranded oligonucleotides to larger plasmid double-stran...

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Autores principales: Priegue, Juan M., Lostalé-Seijo, Irene, Crisan, Daniel, Granja, Juan R., Fernández-Trillo, Francisco, Montenegro, Javier
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2018
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6041776/
https://www.ncbi.nlm.nih.gov/pubmed/29653048
http://dx.doi.org/10.1021/acs.biomac.8b00252
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author Priegue, Juan M.
Lostalé-Seijo, Irene
Crisan, Daniel
Granja, Juan R.
Fernández-Trillo, Francisco
Montenegro, Javier
author_facet Priegue, Juan M.
Lostalé-Seijo, Irene
Crisan, Daniel
Granja, Juan R.
Fernández-Trillo, Francisco
Montenegro, Javier
author_sort Priegue, Juan M.
collection PubMed
description [Image: see text] The recent advances in genetic engineering demand the development of conceptually new methods to prepare and identify efficient vectors for the intracellular delivery of different nucleotide payloads ranging from short single-stranded oligonucleotides to larger plasmid double-stranded circular DNAs. Although many challenges still have to be overcome, polymers hold great potential for intracellular nucleotide delivery and gene therapy. We here develop and apply the postpolymerization modification of polyhydrazide scaffolds, with different degree of polymerization, for the preparation of amphiphilic polymeric vehicles for the intracellular delivery of a circular plasmid DNA. The hydrazone formation reactions with a mixture of cationic and hydrophobic aldehydes proceed in physiologically compatible aqueous conditions, and the resulting amphiphilic polyhydrazones are directly combined with the biological cargo without any purification step. This methodology allowed the preparation of stable polyplexes with a suitable size and zeta potential to achieve an efficient encapsulation and intracellular delivery of the DNA cargo. Simple formulations that performed with efficiencies and cell viabilities comparable to the current gold standard were identified. Furthermore, the internalization mechanism was studied via internalization experiments in the presence of endocytic inhibitors and fluorescence microscopy. The results reported here confirmed that the polyhydrazone functionalization is a suitable strategy for the screening and identification of customized polymeric vehicles for the delivery of different nucleotide cargos.
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spelling pubmed-60417762018-07-15 Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection Priegue, Juan M. Lostalé-Seijo, Irene Crisan, Daniel Granja, Juan R. Fernández-Trillo, Francisco Montenegro, Javier Biomacromolecules [Image: see text] The recent advances in genetic engineering demand the development of conceptually new methods to prepare and identify efficient vectors for the intracellular delivery of different nucleotide payloads ranging from short single-stranded oligonucleotides to larger plasmid double-stranded circular DNAs. Although many challenges still have to be overcome, polymers hold great potential for intracellular nucleotide delivery and gene therapy. We here develop and apply the postpolymerization modification of polyhydrazide scaffolds, with different degree of polymerization, for the preparation of amphiphilic polymeric vehicles for the intracellular delivery of a circular plasmid DNA. The hydrazone formation reactions with a mixture of cationic and hydrophobic aldehydes proceed in physiologically compatible aqueous conditions, and the resulting amphiphilic polyhydrazones are directly combined with the biological cargo without any purification step. This methodology allowed the preparation of stable polyplexes with a suitable size and zeta potential to achieve an efficient encapsulation and intracellular delivery of the DNA cargo. Simple formulations that performed with efficiencies and cell viabilities comparable to the current gold standard were identified. Furthermore, the internalization mechanism was studied via internalization experiments in the presence of endocytic inhibitors and fluorescence microscopy. The results reported here confirmed that the polyhydrazone functionalization is a suitable strategy for the screening and identification of customized polymeric vehicles for the delivery of different nucleotide cargos. American Chemical Society 2018-04-13 2018-07-09 /pmc/articles/PMC6041776/ /pubmed/29653048 http://dx.doi.org/10.1021/acs.biomac.8b00252 Text en Copyright © 2018 American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
spellingShingle Priegue, Juan M.
Lostalé-Seijo, Irene
Crisan, Daniel
Granja, Juan R.
Fernández-Trillo, Francisco
Montenegro, Javier
Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection
title Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection
title_full Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection
title_fullStr Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection
title_full_unstemmed Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection
title_short Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection
title_sort different-length hydrazone activated polymers for plasmid dna condensation and cellular transfection
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6041776/
https://www.ncbi.nlm.nih.gov/pubmed/29653048
http://dx.doi.org/10.1021/acs.biomac.8b00252
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