Optimization of Transfection of Mutated Phosphoinositide-3-Kinase in a Study of Staphylococcus aureus Host Invasion
Kelsey Haaning and Susan A. McDowell*
Ball State University, Muncie, IN, USA
*Corresponding author: firstname.lastname@example.org
We have begun to investigate the role of phosphoinositide-3-kinase (PI3K) in host cell invasion by Staphylococcus aureus, the most common etiologic agent of sepsis .One of the model systems for these invasion studies is the human embryonic kidney (HEK) 293A cell line. These cells
are relevant to the clinical experience of systemic infection and are useful for transient expression of mutated constructs by adenoviral infection or by the use of transfection reagents. We optimized the method for expressing mutated constructs using FuGENE® HD Transfection Reagent or another commercially available transfection reagent (reagent L). We found that HEK 293A cells were less adherent in the presence of reagent L compared with FuGENE® HD Transfection Reagent, resulting in greater variability in the S. aureus invasion assay.