RT Journal Article
T1 Different-Length Hydrazone Activated Polymers for Plasmid DNA Condensation and Cellular Transfection
A1 Priegue Caamaño, Juan Manuel
A1 Lostalé Seijo, Irene
A1 Crisan, Daniel
A1 Granja Guillán, Juan Ramón
A1 Fernández Trillo, Francisco
A1 Montenegro García, Javier
K1 Aldehydes
K1 Genetics
K1 Fluorescence
K1 Gene delivery
K1 Polymers
AB 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
PB American Chemical Society
SN 1525-7797
YR 2018
FD 2018
LK http://hdl.handle.net/10347/24313
UL http://hdl.handle.net/10347/24313
LA eng
NO Biomacromolecules 2018, 19, 7, 2638–2649
NO This work was partially supported by the Spanish Agencia Estatal de Investigación (AEI) [CTQ2014-59646-R, CTQ2016-78423-R, SAF2017-89890-R], the Xunta de Galicia (ED431G/09, ED431C 2017/25 and 2016-AD031), the ERDF, Royal Society U.K (IE130688 and RG140273), and the Wellcome Trust (177ISSFPP). J.M. received a Ramón y Cajal (RYC-2013-13784), an ERC Starting Investigator Grant (DYNAP-677786), and a Young Investigator Grant from the Human Frontier Science Research Program (RGY0066/2017). F. F.-T. thanks the Birmingham Science City and the European Regional Development Fund, and the University of Birmingham (John Evans Fellowship). J.M.P. thanks the Spanish Agencia Estatal de Investigación (AEI) for his research contract
DS Minerva
RD 3 may 2026