Nucleophile responsive charge-reversing polycations for pDNA transfection
Reece W. Lewis (TU Delft - ChemE/Advanced Soft Matter)
A. Muralidharan (TU Delft - BN/Bionanoscience, Kavli institute of nanoscience Delft, TU Delft - BN/Stan Brouns Lab)
B. Klemm (TU Delft - ChemE/Advanced Soft Matter)
PE Boukany (TU Delft - ChemE/Product and Process Engineering)
R Eelkema (TU Delft - ChemE/Advanced Soft Matter)
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Abstract
Polycationic carriers promise low cost and scalable gene therapy treatments, however inefficient intracellular unpacking of the genetic cargo has limited transfection efficiency. Charge-reversing polycations, which transition from cationic to neutral or negative charge, can offer targeted intracellular DNA release. We describe a new class of charge-reversing polycation which undergoes a cationic-to-neutral conversion by a reaction with cellular nucleophiles. The deionization reaction is relatively slow with primary amines, and much faster with thiols. In mammalian cells, the intracellular environment has elevated concentrations of amino acids (∼10×) and the thiol glutathione (∼1000×). We propose this allows for decationization of the polymeric carrier slowly in the extracellular space and then rapidly in the intracellular milleu for DNA release. We demonstrate that in a lipopolyplex formulation this leads to both improved transfection and reduced cytotoxicity when compared to a non-responsive polycationic control.
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