Prolonged durability of electroporation microarrays as a result of addition of saccharides to nucleic acids.
The electroporation microarray is a useful tool for high-throughput analysis of gene functions. However, transfection efficiency is greatly impaired by storage of the microarrays, due to water evaporation from arrayed nucleotides. In this study, we aimed at evaluating the effect of saccharides and sugar alcohols, added to the solution of the plasmid DNA or small interfering RNA (siRNA). Microarrays loaded with plasmids and siRNAs were prepared with various polyols including sugars and sugar alcohols. After storage of these microarrays at different temperatures for various time periods, transfection efficiency was evaluated using human embryonic kidney cells. In the case of plasmid-loaded microarrays, addition of monosaccharides (glucose, fructose), disaccharides (trehalose, sucrose), and trisaccharide (raffinose) served to retain transfection efficiency at a reasonably high level after storage at -20 degrees C. The observed effects may be because moisture retention serves to maintain the solubility of DNA. In contrast, polysaccharide (dextran) and sugar alcohol (glycerol) had insignificant effects on retention of transfection efficiency. On the other hand, addition of saccharides and sugar alcohols had insignificant effects on the transfection of siRNA after storage of a microarray at 25 degrees C for 7 days, presumably due to the intrinsically-high solubility of siRNA which consists of short nucleotides.
Fujimoto
Hiroyuki
H
Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan.
Kato
Koichi
K
Iwata
Hiroo
H
D000327:Adsorption; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry*; D018274:Electroporation / Q000379:methods*; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D009005:Monosaccharides / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000737:chemistry*; D012680:Sensitivity and Specificity; D012995:Solubility; D012996:Solutions / Q000737:chemistry; D013402:Sugar Alcohols / Q000737:chemistry; D013499:Surface Properties; D013696:Temperature; D013997:Time Factors; D014162:Transfection / Q000379:methods
D009005:Monosaccharides; D011134:Polysaccharides; D034741:RNA, Small Interfering; D012996:Solutions; D013402:Sugar Alcohols; D004247:DNA