Gene Transfection Mediated by Catiomers Requires Free Highly Charged Polymer Chains To Overcome Intracellular Barriers

Gene Transfection Mediated by Catiomers Requires Free Highly Charged Polymer Chains To Overcome Intracellular Barriers

Author Albuquerque, Lindomar J. C. Google Scholar
de Castro, Carlos E. Google Scholar
Riske, Karin A. Autor UNIFESP Google Scholar
Carlan da Silva, Maria C. Google Scholar
Muraro, Paulo I. R. Google Scholar
Schmidt, Vanessa Google Scholar
Giacomelli, Cristiano Google Scholar
Giacomelli, Fernando C. Google Scholar
Abstract The prospective use of the block copolymers poly(ethylene oxide)(113)-b-poly[2-(diethylamino)ethyl methacrylate](50) (PEO113-b-PDEA(50)) and poly[oligo(ethylene glycol)methyl ether methacrylate](70)-b-poly[oligo(ethylene glycol)methyl ether methacrylate(10)-co-2-(diethylamino)ethyl methacrylate(47)-co-2-(diisopropylamino) ethyl methacrylate(47)] (POEGMA(70)-b-P(OEGMA(10)-co-DEA(47)-co-DPA(47))) as nonviral gene vectors was evaluated. The polymers are able to properly condense DNA into nanosized particles (R-H approximate to 75 nm), which are marginally cytotoxic and can be uptaken by cells. However, the green fluorescent protein (GFP) expression assays evidenced that DNA delivery is essentially negligible meaning that intracellular trafficking hampers efficient gene release. Subsequently, we demonstrate that cellular uptake and particularly the quantity of GFP-positive cells are substantially enhanced when the block copolymer polyplexes are produced and further supplemented by BPEI chains (branched polyethylenimine). The dynamic light scattering/electrophoretic light scattering/isothermal titration calorimetry data suggest that such a strategy allows the adsorption of BPEI onto the surface of the polyplexes, and this phenomenon is responsible for increasing the size and surface charge of the assemblies. Nevertheless, most of the BPEI chains remain freely diffusing in the systems. The biological assays confirmed that cellular uptake is enhanced in the presence of BPEI and principally, the free highly charged polymer chains play the central role in intracellular trafficking and gene transfection. These investigations pointed out that the transfection efficiency versus cytotoxicity issue can be balanced by a mixture of BPEI and less cytotoxic agents such as for instance the proposed block copolymers.
xmlui.dri2xhtml.METS-1.0.item-coverage Washington
Language English
Sponsor FAPESP
Grant number FAPESP: 2014/22983-9
FAPESP: 2015/24686-4
CNPq: 302467/2014-9
Date 2017
Published in Biomacromolecules. Washington, v. 18, n. 6, p. 1918-1927, 2017.
ISSN 1525-7797 (Sherpa/Romeo, impact factor)
Publisher Amer Chemical Soc
Extent 1918-1927
Access rights Closed access
Type Article
Web of Science ID WOS:000403387100024

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