Gene transfection into adherent cells using electroporation on a dendrimer-modified gold electrode.
Gene transfection into adherent cells from plasmid DNA (pDNA)-arrayed substrates known as gene transfection arrays appears to be a promising tool for the high-throughput analysis of gene functions and protein-protein interaction networks. We tested the ability of electric pulse-stimulated gene transfection from a substrate to overcome low expression efficiency and cross contamination between spots on arrays. We prepared the electrodes used for electric pulse-stimulated gene transfection by sequentially loading a gold thin layer, a self-assembled monolayer of a carboxylic acid-terminated alkanethiol (COOH-SAM), and poly(amidoamine) (PAMAM) dendrimers, either through electrostatic interactions or by covalent linkage to COOH-SAM and then to pDNA. When dendrimers were loaded onto the electrode using electrostatic interactions, the gene-expression efficiency of adherent cells increased as the generation numbers of the dendrimers that we used increased. Gene expression was rarely observed in adherent cells when dendrimers were covalently immobilized onto the electrode. Additionally, we successfully demonstrated site-specific gene transfer using a dendrimer-array electrode with no cross contamination between spots on the electrode.
Koda
S
S
Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Inoue
Y
Y
Iwata
H
H
D000327:Adsorption; D000473:Alkanes / Q000737:chemistry; D002264:Carboxylic Acids / Q000737:chemistry; D002448:Cell Adhesion; D002460:Cell Line; D004247:DNA / Q000235:genetics; D050091:Dendrimers / Q000737:chemistry*; D004566:Electrodes; D018274:Electroporation / Q000295:instrumentation* / Q000379:methods*; D015870:Gene Expression; D006046:Gold / Q000737:chemistry*; D006801:Humans; D010957:Plasmids / Q000235:genetics; D011073:Polyamines / Q000737:chemistry*; D055672:Static Electricity; D013438:Sulfhydryl Compounds / Q000737:chemistry; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013997:Time Factors; D014162:Transfection / Q000295:instrumentation* / Q000379:methods*
D000473:Alkanes; D002264:Carboxylic Acids; D050091:Dendrimers; C531249:Poly(amidoamine); D011073:Polyamines; D013438:Sulfhydryl Compounds; D006046:Gold; D004247:DNA