Peptide-based multicomponent oligonucleotide delivery systems: optimisation of poly-l-lysine dendrons for plasmid DNA delivery

Kamaruzaman, Khairul A., Moyle, Peter M. and Toth, Istvan (2016) Peptide-based multicomponent oligonucleotide delivery systems: optimisation of poly-l-lysine dendrons for plasmid DNA delivery. International Journal of Peptide Research and Therapeutics, 1-16. doi:10.1007/s10989-016-9545-5


Author Kamaruzaman, Khairul A.
Moyle, Peter M.
Toth, Istvan
Title Peptide-based multicomponent oligonucleotide delivery systems: optimisation of poly-l-lysine dendrons for plasmid DNA delivery
Journal name International Journal of Peptide Research and Therapeutics   Check publisher's open access policy
ISSN 1573-3149
1573-3904
Publication date 2016-07-20
Sub-type Article (original research)
DOI 10.1007/s10989-016-9545-5
Open Access Status Not Open Access
Start page 1
End page 16
Total pages 16
Place of publication Dordrecht, Netherlands
Publisher Springer
Collection year 2017
Language eng
Formatted abstract
Gene therapy is a promising means to treat or prevent diseases either through gene silencing or expression. Some of the most effective delivery agents are polycationic dendrimers, which are highly branched constructs incorporating many positively charged groups. Two of the most effective dendrimers are polyethyleneimine (PEI) and poly(amidoamine) (PAMAM), which show high proficiency at overcoming barriers to oligonucleotide delivery. However, because of their abundance of cationic charge, they are associated with severe toxicity. We have therefore aimed to develop a low toxicity oligonucleotide delivery system, incorporating multiple components that have been selected and optimised to overcome the barriers to efficient oligonucleotide delivery. In this work we have focused on improving the toxicity, cellular uptake, and condensation of plasmid DNA (pDNA) through the fusion of synthetic poly-l-lysine (PLL) dendrons with the cell penetrating peptide TAT(48-60). A library of dendron structures, from 4+ to 16+ charge, and constructs containing six histidine residues, were synthesised. The effects of each modification on pDNA binding and condensation; cellular uptake and toxicity; and the size and zeta-potential of the complexes were assessed to identify the optimum dendron for incorporation into our systems. This work concluded that increasing the dendron charge from 4+ to 16+ significantly improved cellular uptake and pDNA condensation, with no effect on toxicity, while PLL dendrons with greater than 16+ charge could not be efficiently produced. In comparison, the incorporation of six histidines into these constructs had a variable effect on cellular uptake, and generated larger sized complexes, but did not affect toxicity.
Keyword Branched poly-l-lysine
Dendron
Histidine
Non-viral gene delivery
Oligonucleotide delivery
Plasmid DNA delivery
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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Created: Sun, 24 Jul 2016, 18:33:47 EST by Peter Moyle on behalf of School of Pharmacy