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Cationic Peptide Mediated Oligonucleotide Delivery to DU145 Prostate Cancer Cells - Cell Surface Binding Detected by High Resolution SEM

Published online by Cambridge University Press:  02 July 2020

Joan F. Karr
Affiliation:
Department of Urology and Department of Chemistry, Emory University, Atlanta, GA30322
Robert P. Apkarian
Affiliation:
Integrated Microscopy & Microanalytical Facility, Department of Chemistry, Emory University, Atlanta, GA30322
John A. Petros
Affiliation:
Department of Urology and Department of Chemistry, Emory University, Atlanta, GA30322
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Extract

Transfection of mammalian cells can be enhanced by forming complexes between exogenous DNA and cationic peptides which facilitate DNA transport through cell membranes. Although complexes of plasmid DNA and cationic peptides have been characterized by microscopy, little is known about the complexes formed between oligonucleotides (ODN) and cationic peptides or how the ultrastructure of these complexes affects the intracellular delivery via the cell membrane. Because ODN have potential as therapeutic reagents in the treatment of many diseases, there is a need to develop improved methods for effective delivery of these molecules. By analyzing molecular interactions between cationic peptides and ODN and their effects on cell membranes, cell viability, and DNA delivery, improved delivery compounds can be designed. In this paper, we compare the interactions of a 28 base ODN with three cationic peptides: a novel compound, MNLEK (H-Met-(Nle-Lys4)7-Nle-NH2), an established transfection reagent, poly(L-lysine) (∼ 54 kDa), and a recently described cationic peptide which delivers both plasmid DNA and antisense ODN to cells, KALA (WEAKLAKALAKALAKHLAKALAKALKACEA).

For high resolution scanning electron microscopy (HRSEM), DU145 human prostate cancer cells were grown overnight on poly-(D-lysine) coated, UV irradiated silicon chips.

Type
Biomaterials
Copyright
Copyright © Microscopy Society of America

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