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AP-4F, antennapedia peptide linked to an amphipathic alpha helical peptide, increases the efficiency of Lipofectamine-mediated gene transfection in endothelial cells. Biochem Biophys Res Commun 2003 Jun 06;305(3):605-10

Date

05/24/2003

Pubmed ID

12763037

DOI

10.1016/s0006-291x(03)00803-9

Scopus ID

2-s2.0-0038239863   15 Citations

Abstract

Typically, endothelial cells are difficult to transfect. In this study, we report that antennapedia peptide (AP) linked to L-4F, a water-soluble, amphipathic alpha helical peptide that avidly binds lipids (AP-4F) increases Lipofectamine 2000-mediated transfection of bovine coronary endothelial cell cultures. Transfection efficiency was monitored by flow cytometry and fluorescent microscopy. Lipofectamine 2000 transfection of endothelial cell cultures with green fluorescence protein (GFP)-DNA typically yields transfection efficiencies of 35.4+/-3.3% with low levels of cell death (8.1+/-1.0%). Pre-treatment of the Lipofectamine 2000-GFP-DNA complexes with AP-4F for 5 min increased transfection to 58.2+/-2.8% without increasing cell death. AP-4F increases Lipofectamine 2000-mediated transfection in a time-dependent fashion (within 10-20 min). Systematic studies reveal that the individual components of AP-4F, i.e., AP and L-4F alone, are ineffective in increasing Lipofectamine 2000-mediated transfection and that AP-4F must be directly associated with DNA liposomes prior to transfection for optimal uptake by endothelial cells. These observations demonstrate that AP-4F may be useful for increasing the transfection efficiency of endothelial cell cultures with standard commercially available reagents.

Author List

Ou J, Geiger T, Ou Z, Ackerman AW, Oldham KT, Pritchard KA Jr

Authors

Keith T. Oldham MD Professor in the Surgery department at Medical College of Wisconsin
Kirkwood A. Pritchard PhD Professor in the Surgery department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Amino Acid Sequence
Animals
Antennapedia Homeodomain Protein
Cation Exchange Resins
Cattle
Cells, Cultured
Endothelium, Vascular
Flow Cytometry
Green Fluorescent Proteins
Homeodomain Proteins
Lipids
Liposomes
Luminescent Proteins
Microscopy, Fluorescence
Molecular Sequence Data
Nuclear Proteins
Peptides
Transcription Factors
Transfection
jenkins-FCD Prod-482 91ad8a360b6da540234915ea01ff80e38bfdb40a