A Novel Cell‐Penetrating Lipopeptide for DNA Binding, Self‐Assembly, and Delivery Into HeLa and Pluripotent Stem Cells
Lucas R. de Mello, Tâmisa Seeko Bandeira Honda, Valeria Castelletto, Patrícia Terra Alves, Sang Won Han, Emerson Rodrigo da Silva, Ian W. Hamley
TL;DR
A new lipopeptide is designed to deliver DNA into HeLa and stem cells, showing better performance than existing methods.
Contribution
A novel cell-penetrating lipopeptide is developed for efficient DNA delivery into pluripotent stem cells.
Findings
The lipopeptide forms micelles and DNA co-assemblies through intercalation.
DNA is successfully delivered into HeLa and mouse embryonic stem cells.
The lipopeptide outperforms lipofectamine in DNA delivery and shows high cytocompatibility.
Abstract
A lipopeptide is designed that contains an epitope from simian virus T‐antigen (SV40T, PKKKRKV) conjugated to an N‐terminal palmitoyl (C16‐) moiety, with the aim to act as an effective cell‐penetrating lipopeptide, with additional aggregation propensity conferred by the lipid chain. A combination of cryo‐TEM and small‐angle X‐ray scattering (SAXS) is used to show that the lipopeptide forms micelles, but mixtures with DNA lead to formation of fractal cluster‐like co‐assemblies due to intercalation of the DNA and peptide. Spectroscopic studies using fluorescence and circular dichroism (along with fiber X‐ray diffraction) show that the peptide interacts with DNA and inserts into the groove. Confocal microscopy along with flow cytometry confirms delivery of DNA into both HeLa and mouse embryonic stem cells (mESCs) in pluripotent state, and the system shows excellent cytocompatibility as…
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Taxonomy
TopicsRNA Interference and Gene Delivery · Lipid Membrane Structure and Behavior · Supramolecular Self-Assembly in Materials