Coupling of importin beta binding peptide on plasmid DNA: transfection efficiency is increased by modification of lipoplexs physico-chemical properties.Report as inadecuate




Coupling of importin beta binding peptide on plasmid DNA: transfection efficiency is increased by modification of lipoplexs physico-chemical properties. - Download this document for free, or read online. Document in PDF available to download.

1 UMR 7001 CNRS - ENSCP - Gencell S.A 2 LPS - Laboratoire Pierre Süe 3 UPCG - Unité de Pharmacologie Chimique et Génétique

Abstract : BACKGROUND: Non-viral vectors for gene transfer are less immunogenic than viral vectors but also less efficient. Significant effort has focused on enhancing non-viral gene transfer efficiency by increasing nuclear import of plasmid DNA, particularly by coupling nuclear localization peptidic sequences to plasmid DNA. RESULTS: We have coupled a 62-aminoacid peptide derived from hSRP1alpha importin beta binding domain, called the IBB peptide to plasmid DNA by using the heterobifunctional linker N-4-azido-2,3,5,6 tetrafluorobenzyl-6-maleimidyl hexanamide TFPAM-6. When covalently coupled to plasmid DNA, IBB peptide did not increase the efficiency of cationic lipid mediated transfection. The IBB peptide was still able to interact with its nuclear import receptor, importin beta, but non-specifically. However, we observed a 20-fold increase in reporter gene expression with plasmid DNA - IBB peptide complexes under conditions of inefficient transfection. In which case, IBB was associated with plasmid DNA through self assembling ionic interaction. CONCLUSIONS: The improvement of transfection activity was not due to an improved nuclear import of DNA, but rather by the modification of physicochemical properties of IBB peptide - plasmid complexes. IBB peptide increased lipoplex size and these larger complexes were more efficient for gene transfer.





Author: Marie Carrière - Virginie Escriou - Aline Savarin - Daniel Scherman -

Source: https://hal.archives-ouvertes.fr/



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