TransMessenger Transfection Reagent

• Highly efficient transfection
• Reproducible results ensured by stringent quality control
• Efficient transfection of primary neuronal cells

TransMessenger Reagent provides a fast and easy procedure and high transfection efficiencies (see figure " TransMessenger Reagent with HeLa cells"). Since the amount of RNA is a critical factor for successful transfection, we recommend optimizing the amounts of RNA and TransMessenger Transfection Reagent for every cell type–RNA combination (see figure " Amount of RNA and TransMessenger Reagent vs. transfection efficiency"). To facilitate this, the reagent is provided with guidelines for optimization together with starting points for optimization in different cell-culture formats.
TransMessenger Transfection Reagent is highly suited for use in functional cardiac cell studies using neonatal rat ventricular cardiac cells.
Cardiomyocytes from cryopreserved, dissociated neonatal rat ventricular cardiac cells (R-CM-561 [QBMCellScience.com]) display excellent viability, pharmacology, morphology, and connectivity, as well as contractile and electrical activity necessary for use in functional screening (see video Transfection control: Functional Cardiomyocytes Beating). To evaluate the suitability of using TransMessenger Transfection Reagent in functional cardiac cell studies, cells were transfected with a GFP-expression vector 4 hours after plating, and were fixed 5 days post-transfection. Use of TransMessenger Transfection Reagent resulted in an excellent, 60% survival rate post-transfection with a transfection efficiency of 10–13%. All cardiomyocytes displayed beating post-transfection (sees videos GPF-Positive Cardiomyocytes Beating Post-Transfection and Functional Cardiomyocytes Beating Post-Transfection), indicating functional cells. The number of transfected cardiomyocyte versus non-cardiomyocyte cells was evaluated and results demonstrate that a high number of cardiomyocyte and non-cardiomyocyte cells were transfected. Equivalent morphologies between transfected and non-transfected cells were also observed (see figure  Cardiac cell studies).

All TransMessenger Reagent components are provided as ready-to-use solutions. To generate TransMessenger–RNA transfection complexes, simply mix your RNA with Enhancer R and Buffer EC-R and incubate for 5 minutes at room temperature, then add TransMessenger Reagent and incubate for a further 5–10 minutes. The complexes are mixed with medium and added directly to the cells. Following a 3 hours incubation, the medium is changed and the cells are incubated until they are ready for analysis.
Optimal transfection results are achieved using high-purity RNA that is free of DNA, proteins, and other contaminants. RNA purified with RNeasy is highly recommended.

Transfection of cells with RNA rather than DNA offers new possibilities for transfection experiments. Transfected RNA sequences are expressed in the absence of transcription, and in a promoter-independent manner. In addition, protein expression usually occurs sooner following transfection of RNA rather than DNA.

RNA transfection with TransMessenger Transfection Reagent can be used for:

• Studies of cells not efficiently transfected with plasmid
• DNA Direct studies of RNA function