Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
1998-2-5
pubmed:abstractText
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.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
1087-2906
pubmed:author
pubmed:issnType
Print
pubmed:volume
7
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
335-43
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:9303185-Amino Acid Sequence, pubmed-meshheading:9303185-Animals, pubmed-meshheading:9303185-Base Sequence, pubmed-meshheading:9303185-COS Cells, pubmed-meshheading:9303185-Cell Membrane Permeability, pubmed-meshheading:9303185-Cell Nucleus, pubmed-meshheading:9303185-Cell Survival, pubmed-meshheading:9303185-Cytosol, pubmed-meshheading:9303185-Hemagglutinin Glycoproteins, Influenza Virus, pubmed-meshheading:9303185-Humans, pubmed-meshheading:9303185-Kinetics, pubmed-meshheading:9303185-Mammals, pubmed-meshheading:9303185-Molecular Sequence Data, pubmed-meshheading:9303185-Oligonucleotides, Antisense, pubmed-meshheading:9303185-Peptides, pubmed-meshheading:9303185-Phosphatidylethanolamines, pubmed-meshheading:9303185-Thionucleotides, pubmed-meshheading:9303185-Tumor Cells, Cultured
pubmed:year
1997
pubmed:articleTitle
Cytosolic and nuclear delivery of oligonucleotides mediated by an amphiphilic anionic peptide.
pubmed:affiliation
Centre de Biophysique Moléculaire, CNRS, Orléans, France.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't