( gents. Both in fetal bovine ...)High-Efficiency Transfections Achieved with New Low-Toxicity Reagent

gents. Both in fetal bovine serum (FBS) and in serum-free environment, GeneJammer-mediated transfection approached an efficiency of 90% to 100% in CHO-K1 and COS-7 cells . Adding FBS increased the amount of transfected HeLa and 293 cells from 38% and 50% to 65% and 92%, respectively. The efficacy of each competitive reagent varied among the different cell lines, with the following proportion of cells transfected: 50 to 95% of CHO-K1 cells, 30 to 80% of COS-7 cells, 10 to 22% of HeLa cells, and 25 to 85% of 293 cells. For all cell lines, transfections with GeneJammer reagent yielded satisfactory efficiencies. Twenty-four hours after transfection, we used Stratagenes in situ b-galactosidase staining kit to stain cells with X-gal, and the resulting cells that expressed for b-galactosidase were photographed (Figure 2).

Fig.2

Assessing Post-transfection Cell Viability

We evaluated the cytotoxic effects of GeneJammer and the competitive reagents by counting the cells stained with trypan blue . Briefly, 24 hours after transfections in the described conditions, media from each well was removed and saved. Cells were trypsinized, combined with saved media, pelleted, and resuspended in 25 l media. Aliquots were combined with an equal volume of 0.4% trypan blue stain. Cells were counted and the ratio of blue to white cells was determined. Of the cells transfected with GeneJammer reagent, 88% to 98% were viable 24 hours posttransfection, whereas the competitive reagents were associated with higher toxicity. The proportion of viable transfected cells was defined as a percent of transfected cells multiplied by the percent of viable cells (Figure 4). On all tested cell
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