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Polymers 2018, 10(4), 379; https://doi.org/10.3390/polym10040379

The Length of Hydrophobic Chain in Amphiphilic Polypeptides Regulates the Efficiency of Gene Delivery

1
School of Material Science & Engineering, Jiangsu University, 301 Xuefu Road, Zhenjiang 212013, China
2
School of Environmental and Chemical Engineering, Jiangsu University of Science and Technology, 2 Mengxi Road, Zhenjiang 212003, China
3
Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, Ann Arbor, MI 48109, USA
*
Authors to whom correspondence should be addressed.
Received: 20 March 2018 / Revised: 28 March 2018 / Accepted: 28 March 2018 / Published: 1 April 2018
(This article belongs to the Special Issue Polypeptide Containing Polymers)
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Abstract

The major challenges of non-viral carriers are low transfection efficiency and high toxicity. To overcome this bottleneck, it is very important to investigate the structure-property-function (transfection efficiency) relationships of polycations. Herein, different length hydrophobic poly(l-leucine) chains in amphiphilic polypeptides were precisely synthesized by α-amino acid N-carboxyanhydrides (NCA) ring-opening polymerization and these biocompatible polypeptides were chosen as a model to further examine the transfection in vitro. These polypeptides were characterized by nuclear magnetic resonance spectroscopy (NMR) and size exclusion chromatography (SEC). Agarose gel electrophoresis (AGE) was employed to validate the ability of DNA condensation and transmission electron microscopy (TEM) was used to observe the assemblies of polyplexes. Cytotoxicity was evaluated in COS-7 cell lines and transfection was performed in normal cell COS-7 and cancer cell Hep G2. The results showed that NCA monomers were prepared and the amphiphilic polypeptides, poly(lysine(CBZ))50-block-poly(l-leucine)10, poly(l-lysine(CBZ))50-block-poly(l-leucine)15, and poly(l-lysine(CBZ))50-block-poly(l-leucine)25, were successfully synthesized with controlled molecular weight and narrow distribution. After deprotection of CBZ, these materials can condense plasmid DNA into 100 nm nanoparticles and the cellular uptake of polyplexes was as fast as 30 min. The transfection data shown these materials had a good transfection efficiency comparing to polyethylenimine (Branched, 25 kDa) while they displayed ignored cytotoxicity. More importantly, we discovered the length of hydrophobic poly(l-leucine) in amphiphilic polypeptides steadily regulates gene delivery efficiency in two kinds of cells ranking poly(l-lysine)50-block-poly(l-leucine)25 > poly(l-lysine)50-block-poly(l-leucine)15 > poly(l-lysine)50-block-poly(l-leucine)10. View Full-Text
Keywords: controlled polymerization; biocompatible polypeptides; gene delivery; transfection efficiency; hydrophobic chain; structure-property-function controlled polymerization; biocompatible polypeptides; gene delivery; transfection efficiency; hydrophobic chain; structure-property-function
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Zhang, Y.; Zhou, Z.; Chen, M. The Length of Hydrophobic Chain in Amphiphilic Polypeptides Regulates the Efficiency of Gene Delivery. Polymers 2018, 10, 379.

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