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Enhanced gene transfection with addition of a cell‐penetrating peptide in substrate‐mediated gene delivery
Author(s) -
Chen Si,
Zhuo RenXi,
Cheng SiXue
Publication year - 2010
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.1488
Subject(s) - transfection , gene delivery , gene expression , dna , gene , microbiology and biotechnology , substrate (aquarium) , hek 293 cells , dna vaccination , genetic enhancement , chemistry , biophysics , biology , biochemistry , plasmid , ecology
Background To overcome extracellular barriers in gene therapy, controlled release technology has been adapted to protect DNA from degradation and maintain elevated DNA concentration. Although cell‐penetrating peptides (CPPs) have been widely utilized to enhance the transfection efficiency of solution‐based transfections, as far as we are aware, the effect of these peptides on substrate‐mediated transfection has not yet been reported. Methods In the present study, the effect of a CPP, KALA, on the substrate‐mediated gene delivery was studied. A fast degrading cholic acid functionalized star poly( DL ‐lactide) was used to fabricate Ca‐P/DNA/KALA co‐precipitates‐deposited films and Ca‐P/DNA/KALA co‐precipitates‐encapsulated films for mediating substrate‐mediated transfection. The in vitro gene transfections in HEK293T and HepG2 cells mediated by the different films were investigated. The effect of KALA content on gene transfection was studied. Results Gene expression of film‐mediated transfections could be significantly enhanced by the addition of KALA at a quite low content with a DNA/KALA ratio of 10 : 1. As a result of the surface erosion mechanism of the functionalized star poly( DL ‐lactide), our substrate‐mediated transfection system could rapidly release Ca‐P/DNA/KALA to mediate efficient gene expression. During the transfection, the degradation of the polymer films could be observed and the degradation did not show any unfavorable effects on gene expression. Conclusions The presence of a CPP could significantly enhance the gene expression of substrate‐mediated transfections. Copyright © 2010 John Wiley & Sons, Ltd.