DNA transfection into HEK-293 cell line using EcoTransfect

HEK-293 cell line was transiently and stably transfected using EcoTransfect reagent from OZ Biosciences

This article illustrates the efficiency of EcoTransfect transfection reagent from OZ Biosciences to stably and transiently genetically modify HEK-293 cells.

article reference: Cell Signal. 2013 Nov 21. pii: S0898-6568(13)00352-5. 

β-Catenin-dependent pathway activation by both promiscuous "canonical" WNT3a-, and specific "noncanonical" WNT4- and WNT5a-FZD receptor combinations with strong differences in LRP5 and LRP6 dependency.

Ring L, Neth P, Weber C, Steffens S, Faussner A.

AbstractThe WNT/β-catenin signalling cascade is the best-investigated frizzled receptor (FZD) pathway, however, whether and how specific combinations of WNT/FZD and co-receptors LRP5 and LRP6 differentially affect this pathway are not well understood. This is mostly due to the fact that there are 19 WNTs, 10 FZDs and at least two co-receptors. In our attempt to identify the signalling capabilities of specific WNT/FZD/LRP combinations we made use of our previously reported TCF/LEF Gaussia luciferase reporter gene HEK293 cell line (Ring et al., 2011). Generation of WNT/FZD fusion constructs - but not their separate transfection - without or with additional isogenic overexpression of LRP5 and LRP6 in our reporter cells permitted the investigation of specific WNT/FZD/LRP combinations. The canonical WNT3a in fusion to almost all FZDs was able to induce β-catenin-dependent signalling with strong dependency on LRP6 but not LRP5. Interestingly, noncanonical WNT ligands, WNT4 and WNT5a, were also able to act "canonically" but only in fusion with specific FZDs and with selective dependence on LRP5 or LRP6. These data and extension of this experimental setup to the poorly characterized other WNTs should facilitate deeper insight into the complex WNT/FZD signalling system and its function.

EcoTransfect from OZ Biosciences is an economical reagent dedicated to the transfection of popular cell lines. It is the perfect solution to quickly analyze the biological activity of your nucleic acids, to perform routine transfection assays at low cost and to accomplish high throughput screening.