title: Synthesis and characterization of polyaspartamide copolymers obtained by ATRP for nucleic acid delivery.
abstract: Nucleic acid molecules such as small interfering RNAs (siRNAs) and plasmidic DNAs (pDNAs) have been shown to have the potential to be of therapeutic value in different human diseases. Their practical use is however compromised by the lack of appropriate release systems. Delivered as naked molecules, siRNAs/pDNAs are rapidly degraded by extracellular nucleases thus considerably reducing the amount of molecule which can reach the target cells. Additionally, the anionic charge of the phosphate groups present on the siRNAs/pDNAs backbone, disfavors the interaction with the negatively charged surface of the cell membrane. In this paper we describe the generation of a novel polymer able to deliver both siRNAs and pDNAs. The combined release of these molecules is used in many different experimental settings such as the evaluation of the silencing efficiency of a given siRNA targeted against a given RNA, encoded by the pDNA. The possibility to use the same delivery system is very convenient from the technical point of view and it allows minimizing possible artifacts introduced by the use of different delivery agents for siRNAs and pDNA. The copolymer described here is based on α,β-poly(N-2-hydroxyethyl)-d,l-aspartamide (PHEA) bearing positively chargeable side oligochains, with diethylamino ethyl methacrylate (DEAEMA) as monomer. Monomer polymerization has been obtained by atom transfer radical polymerization (ATRP), a technique which allows the precise polymerization of the monomer. In addition to the chemical-physical characterization of the polymer, we provide evidences of the polymer ability to delivery both siRNAs and pDNA to cultured cells. Whereas additional investigations are necessary to study the delivery mechanisms of this polyplex, the polymer generated represents a novel and convenient device for the delivery of both siRNAs and pDNA.
authors.affiliation: Lab of Biocompatible Polymers, Dipartimento di Scienze e Tecnologie Biologiche, Chimiche, Farmaceutiche (STEBICEF), University of Palermo, via Archirafi 32, Palermo 90123, Italy. Electronic address: gennara.cavallaro@unipa.it.
authors.forename: G
authors.initials: G
authors.lastname: Cavallaro
authors.affiliation: Lab of Biocompatible Polymers, Dipartimento di Scienze e Tecnologie Biologiche, Chimiche, Farmaceutiche (STEBICEF), University of Palermo, via Archirafi 32, Palermo 90123, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Licciardi
authors.affiliation: Lab of Biocompatible Polymers, Dipartimento di Scienze e Tecnologie Biologiche, Chimiche, Farmaceutiche (STEBICEF), University of Palermo, via Archirafi 32, Palermo 90123, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Amato
authors.affiliation: Lab of Biocompatible Polymers, Dipartimento di Scienze e Tecnologie Biologiche, Chimiche, Farmaceutiche (STEBICEF), University of Palermo, via Archirafi 32, Palermo 90123, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Sardo
authors.affiliation: Lab of Biocompatible Polymers, Dipartimento di Scienze e Tecnologie Biologiche, Chimiche, Farmaceutiche (STEBICEF), University of Palermo, via Archirafi 32, Palermo 90123, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Giammona
authors.affiliation: Department of Engineering and Architecture, University of Trieste, Via Alfonso Valerio, 6/A, Trieste I-34127, Italy.
authors.forename: R
authors.initials: R
authors.lastname: Farra
authors.affiliation: Department of Life Sciences, Cattinara University Hospital, Trieste University, Strada di Fiume 447, Trieste I-34149, Italy.
authors.forename: B
authors.initials: B
authors.lastname: Dapas
authors.affiliation: Department of Engineering and Architecture, University of Trieste, Via Alfonso Valerio, 6/A, Trieste I-34127, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Grassi
authors.affiliation: Department of Life Sciences, Cattinara University Hospital, Trieste University, Strada di Fiume 447, Trieste I-34149, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Grassi
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry*; D050687:E2F1 Transcription Factor / Q000235:genetics; D006801:Humans; D008689:Methacrylates / Q000737:chemistry*; D051379:Mice; D010455:Peptides / Q000737:chemistry*; D010957:Plasmids; D058105:Polymerization; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000737:chemistry*; D014162:Transfection / Q000379:methods
chemical_list: D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; D008689:Methacrylates; D010455:Peptides; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C008965:alpha,beta-poly((2-hydroxyethyl)-aspartamide); D004247:DNA; C519537:diethylaminoethyl methacrylate
keywords: ATRP; DNA delivery; Polyaspartamide; SiRNA delivery