The story of ViaFect begins with Promega Custom Assay Services (CAS), a group that uses Promega technologies to construct made-to-order assays, typically in a cell line. Many projects from the CAS group involve transfecting cells with expression vectors and reporter vectors. In some instances, customers contact CAS to have an assay constructed in a difficult cell line, after attempting and failing, or experiencing difficulty building the assay themselves.
CAS projects start with a proof-of-concept experiment using transient transfection before moving on to production of a clonal, stable cell line. For difficult cell lines, the CAS group previously turned to electroporation after exhausting lipid-based transfection options. Electroporation often worked, but success came with a price—cytotoxicity. The CAS group challenged R&D to find a better solution—better transfection with low toxicity for difficult-to-use cells. The result of that challenge is the ViaFect™ Transfection Reagent.
The figure demonstrates how ViaFect™ Transfection Reagent helped confirmed the concept, with a usable transient transfection prior to the selection of stable TF-1 suspension cells. On the other hand, as shown, electroporation did not result in significant transfection of the cells.
Figure 1. ViaFect™ Transfection Reagent enables an assay for cytokine signaling in a hematopoietic cell model. TF-1 suspension cells were transiently transfected with pGL4.32[luc2P/NF-κB-RE/Hygro] Vector, an NF-κB response element luciferase reporter, using either ViaFect™ Transfection Reagent at a 2:1 reagent:DNA ratio or Amaxa Nucleofector® II (electroporation). The following day cells were stimulated with TNFα for 6 hours, and the response was measured with Bio-Glo™ Luciferase Reagent.
Primary cells are often difficult to transfect due to their slow growth and high rate of death. ViaFect™ Reagent can be the perfect solution for such cells. These peer-reviewed articles demonstrate the use of ViaFect™ Reagent in the transfection of primary cells:
||Straka, E. et al. (2016) Mercury toxicokinetics of the healthy human term placenta involve amino acid transporters and ABC transporters. Toxicology 340, 34–42.
|Mouse Cortical Neurons
||Egusa, S.F. et al. (2016) Classic cadherin expressions balance postnatal neuronal positioning and dendrite dynamics to elaborate the specific cytoarchitecture of the mouse cortical area. Neurosci. Res. 105, 49–64.
|Mouse Embryonic Fibroblasts
||Peng, Y. et al. (2016) AGE-RAGE signal generates a specific NF-κB RelA “barcode” that directs collagen I expression. Sci. Rep. 6, 18822.
|Mouse Bone Marrow Macrophages
||Naujoks, J. et al. (2016) IFNs modify the proteome of Legionella-containing vacuoles and restrict infection via IRG1-derived itaconic acid. PLoS Pathogens 12, e1005408.
|Mouse Alveolar Macrophages
|Rat Aortic Smooth Muscle Cells
||Horita, H. et al. (2016) Nuclear PTEN functions as an essential regulator of SRF-dependent transcription to control smooth muscle differentiation. Nat. Comm. 7, 10830.
|Human Renal Proximal Tubule Epithelial Cells
||Bethge, T. et al. (2015) Sp1 sites in the noncoding control region of BK polyomavirus are key regulators of bidirectional viral early and late gene expression. J. Virol. 89, 3396–411.
|Mouse Male Germline Stem Cells
||Huang, Y. et al. (2015) Specific tandem 3’UTR patterns and gene expression profiles in mouse Thy1+ germline stem cells. PLoS One 10, e0145417.
|Mouse Endothelial Fibroblasts
||Takahasi, M. et al. (2015) Normalization of overexpressed α-synuclein causing Parkinson’s disease by a moderate gene silencing with RNA interference. Mol. Ther. Nucleic Acids 12, e241.
|Human Umbilical Vein Endothelial Cells
||Yokota, Y. et al. (2015) Endothelial Ca2+ oscillations reflect VEGFR signaling-regulated angiogenic capacity in vivo. eLife 4, e08817.
||Nakmura, T. et al. (2014) Epiprofin orchestrates epidermal keratinocyte proliferation and differentiation. J. Cell Sci. 127, 5261–72.
|The following two tabs change content below.
A former Technical Services Scientist, Kyle has also worked with R&D for product development and now specializes in supporting Promega cellular analysis products in North America.