A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency

Aristides D Tagalakis,1,* Sara Castellaro,1,2,* Haiyan Zhou,1 Alison Bienemann,3 Mustafa M Munye,1 David McCarthy,4 Edward A White,3 Stephen L Hart1 1Experimental and Personalised Medicine Section, University College London (UCL) Institute of Child Health, London, UK; 2Department of Pharmacy, Univ...

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Autores principales:	Tagalakis AD, Castellaro S, Zhou H, Bienemann A, Munye MM, McCarthy D, White EA, Hart SL
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2015
Materias:	Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/e507afb0fc094f9b8e4391e87909d112
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spelling	oai:doaj.org-article:e507afb0fc094f9b8e4391e87909d1122021-12-02T03:56:41ZA method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency1178-2013https://doaj.org/article/e507afb0fc094f9b8e4391e87909d1122015-04-01T00:00:00Zhttp://www.dovepress.com/a-method-for-concentrating-lipid-peptide-dna-and-sirna-nanocomplexes-t-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013 Aristides D Tagalakis,1,* Sara Castellaro,1,2,* Haiyan Zhou,1 Alison Bienemann,3 Mustafa M Munye,1 David McCarthy,4 Edward A White,3 Stephen L Hart1 1Experimental and Personalised Medicine Section, University College London (UCL) Institute of Child Health, London, UK; 2Department of Pharmacy, University of Genova, Genova, Italy; 3Functional Neurosurgery Research Group, School of Clinical Sciences, AMBI Labs, University of Bristol, Southmead Hospital, Bristol, UK; 4UCL School of Pharmacy, London, UK *These authors contributed equally to this work Abstract: Nonviral gene and small interfering RNA (siRNA) delivery formulations are extensively used for biological and therapeutic research in cell culture experiments, but less so in in vivo and clinical research. Difficulties with formulating the nanoparticles for uniformity and stability at concentrations required for in vivo and clinical use are limiting their progression in these areas. Here, we report a simple but effective method of formulating monodisperse nanocomplexes from a ternary formulation of lipids, targeting peptides, and nucleic acids at a low starting concentration of 0.2 mg/mL of DNA, and we then increase their concentration up to 4.5 mg/mL by reverse dialysis against a concentrated polymer solution at room temperature. The nanocomplexes did not aggregate and they had maintained their biophysical properties, but, importantly, they also mediated DNA transfection and siRNA silencing in cultured cells. Moreover, concentrated anionic nanocomplexes administered by convection-enhanced delivery in the striatum showed efficient silencing of the ß-secretase gene BACE1. This method of preparing nanocomplexes could probably be used to concentrate other nonviral formulations and may enable more widespread use of nanoparticles in vivo. Keywords: nanoparticles, concentration, anionic liposome, siRNA, DNA, targeted gene deliveryTagalakis ADCastellaro SZhou HBienemann AMunye MMMcCarthy DWhite EAHart SLDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2015, Iss default, Pp 2673-2683 (2015)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920
spellingShingle	Medicine (General) R5-920 Tagalakis AD Castellaro S Zhou H Bienemann A Munye MM McCarthy D White EA Hart SL A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency
description	Aristides D Tagalakis,1,* Sara Castellaro,1,2,* Haiyan Zhou,1 Alison Bienemann,3 Mustafa M Munye,1 David McCarthy,4 Edward A White,3 Stephen L Hart1 1Experimental and Personalised Medicine Section, University College London (UCL) Institute of Child Health, London, UK; 2Department of Pharmacy, University of Genova, Genova, Italy; 3Functional Neurosurgery Research Group, School of Clinical Sciences, AMBI Labs, University of Bristol, Southmead Hospital, Bristol, UK; 4UCL School of Pharmacy, London, UK *These authors contributed equally to this work Abstract: Nonviral gene and small interfering RNA (siRNA) delivery formulations are extensively used for biological and therapeutic research in cell culture experiments, but less so in in vivo and clinical research. Difficulties with formulating the nanoparticles for uniformity and stability at concentrations required for in vivo and clinical use are limiting their progression in these areas. Here, we report a simple but effective method of formulating monodisperse nanocomplexes from a ternary formulation of lipids, targeting peptides, and nucleic acids at a low starting concentration of 0.2 mg/mL of DNA, and we then increase their concentration up to 4.5 mg/mL by reverse dialysis against a concentrated polymer solution at room temperature. The nanocomplexes did not aggregate and they had maintained their biophysical properties, but, importantly, they also mediated DNA transfection and siRNA silencing in cultured cells. Moreover, concentrated anionic nanocomplexes administered by convection-enhanced delivery in the striatum showed efficient silencing of the ß-secretase gene BACE1. This method of preparing nanocomplexes could probably be used to concentrate other nonviral formulations and may enable more widespread use of nanoparticles in vivo. Keywords: nanoparticles, concentration, anionic liposome, siRNA, DNA, targeted gene delivery
format	article
author	Tagalakis AD Castellaro S Zhou H Bienemann A Munye MM McCarthy D White EA Hart SL
author_facet	Tagalakis AD Castellaro S Zhou H Bienemann A Munye MM McCarthy D White EA Hart SL
author_sort	Tagalakis AD
title	A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency
title_short	A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency
title_full	A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency
title_fullStr	A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency
title_full_unstemmed	A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency
title_sort	method for concentrating lipid peptide dna and sirna nanocomplexes that retains their structure and transfection efficiency
publisher	Dove Medical Press
publishDate	2015
url	https://doaj.org/article/e507afb0fc094f9b8e4391e87909d112
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A method for concentrating lipid peptide DNA and siRNA nanocomplexes that retains their structure and transfection efficiency

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