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Enhancement of gene transfer using YIGSR analog of Tat‐derived peptide
Author(s) -
Saleh Amer F. Alhaj,
Aojula Harmesh S.,
Pluen Alain
Publication year - 2008
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.20854
Subject(s) - ht1080 , transfection , chemistry , reporter gene , microbiology and biotechnology , peptide , flow cytometry , gene , dna , cell culture , biochemistry , gene expression , biology , genetics
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. Biopolymers 89: 62–71, 2008. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com