Cytosolic and Nuclear Delivery of Oligonucleotides Mediated by an Amphiphilic Anionic Peptide

Abstract : Antisense oligonucleotides (ODN) were easily introduced into the cytosol of mammalian cells on permeabilization of the plasma membrane by an amphiphilic anionic peptide. The E5CA peptide (GLFEAIAEFIEGGWEGLIEGCA) is an E5 peptide analog derived from the N-terminal segment of the HA2 subunit of influenza virus hemagglutinin. This peptide undergoes a conformational change when the pH shifts from neutral to around 6.0, inducing a transient permeabilization of the plasma membrane. In the presence of the E5CA peptide at pH close to 6.0, fluoresceinylated ODN were rapidly taken up by cells and diffused into the nucleus. The uptake of ODN was dependent on the E5CA peptide concentration and on the duration of the incubation at low pH, as shown by confocal microscopy and flow cytometry analyses. This procedure is suitable for loading adherent cells as well as nonadherent cells with single-stranded or double-stranded ODN. Under optimal conditions, a high percentage of cells were nuclei loaded, and the viability was not affected. This method makes use of a well-defined chemical product without the requirement of any special equipment. It will be useful to study the interactions of single-stranded or double-stranded ODN used as antisense, antigenes, or decoys.
Document type :
Journal articles
Complete list of metadatas

https://hal.archives-ouvertes.fr/hal-02136009
Contributor : Laëtitia Legoupil <>
Submitted on : Tuesday, May 21, 2019 - 4:55:25 PM
Last modification on : Thursday, June 20, 2019 - 12:22:09 PM

Identifiers

Collections

Citation

Chantal Pichon, Isabelle Freulon, Patrick Midoux, Roger Mayer, Michel Monsigny, et al.. Cytosolic and Nuclear Delivery of Oligonucleotides Mediated by an Amphiphilic Anionic Peptide. Antisense and Nucleic Acid Drug Development, Mary Ann Liebert, 1997, 7 (4), pp.335-343. ⟨10.1089/oli.1.1997.7.335⟩. ⟨hal-02136009⟩

Share

Metrics

Record views

8