Delivery of oligonucleotides into mammalian cells by anionic peptides: comparison between monomeric and dimeric peptides

Abstract

The use of antisense oligonucleotides as putative therapeutic agents is limited by their poor delivery into the cytosol and/or the nucleus because they are not able to efficiently cross lipid bilayers. To circumvent this pitfall, anionic amphipathic peptides derived from the influenza virus fusogenic peptide have been used to destabilize membranes in an acidic environment. In this paper, we compare the ability of a monomeric and a dimeric peptide to introduce oligonucleotides into the cytosol and nuclei of several types of cultured cells. Cells incubated at pH 6.2 or at a slightly lower pH in the presence of the monomeric peptide but not the dimeric peptide were efficiently permeabilized. The location of fluorescent derivatives of peptides and of oligonucleotides was assessed by confocal microscopy. Both the peptides and oligonucleotides remained entrapped in vesicular compartments at neutral pH; at acidic pH, oligonucleotides in the presence of the monomeric peptide were mainly in the nucleus, while in the presence of the dimeric peptide they co-localized with the peptide into vesicles. The data are interpreted on the basis of the spectroscopic behaviour of monomeric and dimeric peptides in relation to the environmental pH.

Carboxylated Superparamagnetic Iron Oxide Particles Label Cells Intracellularly Without Transfection Agents

“…Therefore, we hypothesize that the negatively charged carboxyl groups of the carboxydextran are responsible for the differences seen between Resovist \ and Feridex \ . Although most transfection agents are cationic, i.e., positively charged, also negatively charged substances can exert such a function [31,32]. These anionic transfection agents have been mostly used to transfect small molecules or DNA but not nanoparticles.…”

Delivery of Macromolecules into Live Cells by Simple Co‐incubation with a Peptide

“…osmotic lysis of pinocytic vesicles, plasma membrane permeabilization, scrape loading, or microinjection) and therefore represents a step towards making the delivery of macromolecules to cultured cells a routine procedure for imaging or cell biology applications. [21–23]…”

Modification of Alternative Splicing by Antisense Oligonucleotides as a Potential Chemotherapy for Cancer and Other Diseases

“…Although differences between carriers were found, none seemed to offer a quantum leap in effectiveness and utility. Additional agents, tested in non-splicing assays, included dendrimers [169,170], protamine nanoparticles [171][172][173], cationic porphyrins [174] and anionic peptides [175]. These methods show promise in initial studies but require further investigation in order to determine their potential therapeutic value.…”