Superior transfection efficiencies ranging between 70 and 90% were obtained when using jetPRIME™ reagent versus the top competitor’s reagent for several commonly used cell lines. (Fig. 1).

Fig. 1. Transfection efficiency assessed by FACS analysis in various cell lines 24 h following transfection in 24-well plates according to the manufacturer’s recommendation for competitor L2K  and 0.5 µg plasmid, 1 µl reagent per well for jetPRIME™.

Many other cell lines of various origins, as well as primary cells, are transfected with unusually high percentages (Table 1).

Table 1. Transfection efficiency of various cell types using jetPRIME. The percentage of GFP-positive cells was determined by FACS analysis 24 h after transfection.

jetPRIME™ is such a powerful in vitro transfection reagent that only requires a small amount of reagent and plasmid DNA (Table 2), making it very economical.

Table 2. Amounts of DNA and reagent (jetPRIME™ and competitor) added per well in 6-well plate for transfection according to manufacturers’ recommendations.

In addition to reducing costs, using less DNA also minimizes adverse cytotoxic effects triggered by transfection. Hence, jetPRIME™ is the reagent of choice for high transfection efficiency with excellent cell viability.

jetPRIME™ is extremely gentle to cells during transfection leading to increased cell viability (Fig. 2) and improved transfection results. Cells transfected with jetPRIME™ are healthy, while major cytotoxicity is observed with competitor.

Fig. 2. Phase contrast microscopy of HeLa cells 24 h after transfections performed according to the manufacturer’s recommendations for each reagent.

jetPRIME™ leads to over 90% knockdown of endogenous gene expression in a variety of cell lines. For example, jetPRIME™-mediated transfection of HeLa cells with 10 nM siRNA duplexes targeting endogenous lamin A/C in HeLa cells drastically reduces lamin A/C gene expression to barely detectable level (Fig. 3).

Fig. 3. Endogenous lamin A/C silencing using jetPRIME™. HeLa cells were transfected with 10 nM of 21-mer lamin A/C siRNA. After 48 h, lamin A/C silencing was assessed by immunofluorescence microscopy using an antibody against lamin A/C.

jetPRIME™ is well suited for DNA and siRNA cotransfection experiments. It shows highly efficient gene silencing in a variety of cell lines with very low toxicity. Over 90% silencing is achieved in adherent cells, using 10 nM siRNA (Fig. 4).

In mammalian cells, RNA interference can be achieved by other means than synthetic siRNA. Another approach consists in using plasmid-based methods. It has been reported that efficient gene silencing can be obtained with small hairpin RNA (shRNA) plasmids. For transfection of such plasmids, jetPRIME™, our versatile DNA transfection reagent, is recommended.

When studying miRNA (micro-RNA) using a plasmid-based approach, the standard transfection method using jetPRIME™ is applicable and ensures reliable transfection efficiency.

jetPRIME™ is an easy-to-use transfection reagent (Fig. 5):

• Fast and easy to scale up and down

• Compatible with serum and antibiotics