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Effects of trehalose polycation end-group functionalization on plasmid DNA uptake and transfection
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Anderson, Kevin, Sizovs, Antons, Cortez, Mallory, Waldron, Christopher, Haddleton, David M. and Reineke, Theresa M. (2012) Effects of trehalose polycation end-group functionalization on plasmid DNA uptake and transfection. Biomacromolecules, Volume 13 (Number 8). pp. 2229-2239. doi:10.1021/bm300471n ISSN 1525-7797.
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Official URL: http://dx.doi.org/10.1021/bm300471n
Abstract
In this study, we have synthesized six analogs of a trehalose- pentaethylenehexamine glycopolymer (Tr4) that contain (1A) adamantane, (1B) carboxy, (1C) alkynyl-oligoethyleneamine, (1D) azido trehalose, (1E) octyl, or (1F) oligoethyleneamine end groups and evaluated the effects of polymer end group chemistry on the ability of these systems to bind, compact, and deliver pDNA to cultured HeLa cells. The polymers were synthesized in one-pot azide-alkyne cycloaddition reactions with an adaptation of the Carothers equation for step-growth polymerization to produce a series of polymers with similar degrees of polymerization. An excess of end-capping monomer was added at the end of the polymerizations to maximize functionalization efficiency, which was evaluated with GPC, NMR, and MALDI-TOF. The polymers were all found to bind and compact pDNA at similarly low N/P ratios and form polyplexes with plasmid DNA. The effects of the different end group structures were most evident in the polyplex internalization and transfection assays in the presence of serum as determined by flow cytometry and luciferase gene expression, respectively. The Tr4 polymers end-capped with carboxyl groups (1B) (N/P = 7), octyne (1E) (N/P = 7), and oligoethyleneamine (1F) (N/P = 7), were taken into cells as polyplex and exhibited the highest levels of fluorescence, resulting from labeled plasmid. Similarly, the polymers end-functionalized with carboxyl groups (1E at N/P = 7), octyl groups (1E at N/P = 15), and in particular oligoethyleneamine groups (1F at N/P = 15) yielded dramatically higher reporter gene expression in the presence of serum. This study yields insight into how very subtle structural changes in polymer chemistry, such as end groups can yield very significant differences in the biological delivery efficiency and transgene expression of polymers used for pDNA delivery.
Item Type: | Journal Article | ||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | ||||
Journal or Publication Title: | Biomacromolecules | ||||
Publisher: | American Chemical Society | ||||
ISSN: | 1525-7797 | ||||
Official Date: | 13 August 2012 | ||||
Dates: |
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Volume: | Volume 13 | ||||
Number: | Number 8 | ||||
Page Range: | pp. 2229-2239 | ||||
DOI: | 10.1021/bm300471n | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Restricted or Subscription Access | ||||
Funder: | NIH Director’s New Innovator Award Program | ||||
Grant number: | 1-DP2-OD006669-01 |
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