A peptide nucleic acid-nuclear localization signal fusion that mediates nuclear transport of DNA.
We have combined a peptide nucleic acid (PNA) with the SV40 core nuclear localization signal (NLS), to create a bifunctional PNA-NLS peptide. The PNA-NLS peptide increased the nuclear uptake of oligonucleotides and enhanced the transfection efficacy of plasmids. Gene expression from an enhanced green fluorescent protein plasmid and a lacZ plasmid was preserved when hybridized to PNA-NLS. In combination with the transfection agent polyethyleneimine, we have improved both the nuclear translocation of fluorescence-marked oligonucleotides, and the efficacy of plasmid transfection, up to eightfold. The technique obviates the use of cumbersome coupling procedures of the vector due to DNA-PNA duplex formation or displacement of the antisense plasmid DNA strand by a PNA molecule.
Brandén
L J
LJ
Center for BioTechnology, Department of Biosciences, Karolinska Institutet, NOVUM, SE-14157, Huddinge, Sweden. lars.branden@cbt.ki.se
Mohamed
A J
AJ
Smith
C I
CI
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D019556:COS Cells; D002467:Cell Nucleus / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D005456:Fluorescent Dyes; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D019913:Nuclear Localization Signals; D020135:Peptide Nucleic Acids / Q000378:metabolism*; D010957:Plasmids
D005456:Fluorescent Dyes; D019913:Nuclear Localization Signals; D020135:Peptide Nucleic Acids; D004247:DNA