Enhancement of gene transfer using YIGSR analog of Tat-derived peptide

Amer F Alhaj Saleh, Harmesh S. Aojula, Alain Pluen

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Cell penetrating peptide based gene carriers are notably known for low level of gene transfer. To remedy this, as laminin receptor (LR) has been previously linked to tumor metastasis, the LR-binding domain (YIGSR) as well as a scrambled sequence (SGIYR) were added to Tat-derived peptide sequence (YIGSR-Tat and SGIYR-Tat respectively). Peptides cellular uptake was assessed with high-LR (HT1080) and low-LR (HT29) cell lines by flow cytometry. Their ability to form complexes with DNA was examined using YOPRO-1 fluorescence assay and their transfection efficiencies evaluated using a luciferase reporter gene assay. DNA complexes were formed at (+/-) charge ratios as low as 2:1. While no conclusion could be drawn on the effect of YIGSR sequence on peptides uptake in both cell lines, a significant improvement in gene transfection in HT1080 cells was achieved using YIGSR-Tat compared to Tat and SGIYR-Tat. Additionally this increased efficiency was inhibited by excess free YIGSR. No significant difference in transfection efficiency was observed between Tat, SGIYR-Tat and YIGSR-Tat based complexes in HT29 cells. These studies demonstrate that attachment of receptor-binding ligand (YIGSR) to Tat-derived peptide can improve the efficiency of gene transfer in LR-positive cells (HT1080). © 2007 Wiley Periodicals, Inc.
    Original languageEnglish
    Pages (from-to)62-71
    Number of pages9
    JournalBiopolymers
    Volume89
    Issue number1
    DOIs
    Publication statusPublished - Jan 2008

    Keywords

    • DNA delivery
    • Laminin receptor
    • Targeting
    • Tat peptide
    • YIGSR peptide

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