Reagents optimized for your specific application
Dharmacon transfection reagents are designed specifically for small RNA transfection (DharmaFECT), co-transfection of plasmid and small RNA (DharmaFECT Duo), plasmid transfection (DharmaFECT kb) and self delivering siRNA for difficult to transfect cell types (Accell siRNA).
One of the key success factors in any gene modulation experiment (RNAi, overexpression, gene editing) is the introduction of RNA and/or DNA components into your cell line or in vivo system. When applying lipid-based transfection reagents, there are three general measures of an effective transfection reagent:
We offer a variety of Dharmacon DharmaFECT transfection reagents with demonstrated success for all of these key criteria. Not only do we specialize in transfection of small RNA (siRNA, microRNA, crRNA, tracrRNA) but we offer DharmaFECT Duo, specifically designed for co-delivery of small RNA with plasmid DNA, and DharmaFECT kb for delivery of plasmid DNA.
The most broadly applicable DharmaFECT formulation for optimal siRNA or microRNA transfection into a wide range of cell types for successful RNAi experiments. DharmaFECT 1 has been validated in over 35 cell types.
One of four siRNA/microRNA specific formulations, DharmaFECT 2 is a chemically distinct alternative to one-size-fits-all transfection reagents to achieve high-efficiency silencing in more cell types.
One of four siRNA/microRNA specific formulations, DharmaFECT 3 is a chemically distinct alternative to one-size-fits-all transfection reagents to achieve high-efficiency silencing in more cell types.
DharmaFECT 4 Transfection Reagent is chemically distinct from the other DharmaFECT formulations, providing an alternative when DharmaFECT 1 does not achieve optimal results, and is ideal to include in transfection optimization experiments prior to RNAi screening or other high-value experiments
An aliquot of each of the four DharmaFECT formulations for siRNA/microRNA transfection optimization studies. Useful for determination of the best reagent for your cells and particular experimental conditions.
For high-confidence co-transfection
Efficient transfection of plasmid DNA with minimal cytotoxicity
Targeted gene silencing in difficult-to-transfect cells
The table below lists transfection recommendations to help you select the appropriate DharmaFECT formulation for your research. You can also download a PDF version of the DharmaFECT Cell Type Guide here.
U2OS-(Ubi)EGFP cells were plated at 10,000 cells/well in 96-well plates and co-transfected using DharmaFECT transfection reagents with 25 nM Edit-R Cas9 Nuclease protein NLS and synthetic crRNA:tracrRNA at 50 or 100 nM targeting PSMD7. Cells were harvested 72 hours post-transfection and the relative frequency of gene editing was calculated based on a DNA mismatch detection assay with T7 Endonuclease I. UT = untreated sample, MW = FastRuler Low Range DNA Ladder (Thermo Scientific).
HeLa293T cells were plated at 20,000 cells/well in 96-well plates and co-transfected using DharmaFECT transfection reagents with 200 ng of Edit-R-Cas9 mRNA and synthetic crRNA:tracrRNA targeting VEGFA. Cells were harvested 72 hours post-transfection and the relative frequency of gene editing was calculated based on a DNA mismatch detection assay with T7 Endonuclease I. DF1 = DharmaFECT 1, Duo = DharmaFECT Duo, UT = untreated sample, MW = FastRuler Low Range DNA Ladder (Thermo Scientific).
DharmaFECT reagents are effective across a broader range of experimental conditions when compared to Lipofectamine® 2000 (Invitrogen). Several cell densities and lipid volumes were investigated to determine optimal transfection conditions, shown by the shaded boxes. Three cell densities of HepG2 cells were transfected with GAPD siRNA (100nM) using a range of volumes (0.05 to 1.6μL/well) of Lipofectamine 2000 and DharmaFECT 4 transfection reagents. mRNA levels (bars) were assessed by branched DNA assay (Panomics Quantigene® Reagent System) and cell viability (data points) was determined by alamarBlue® (Biosource International).
To determine transfection efficiencies at low siRNA concentrations, two genes were targeted with various amounts of SMARTpool siRNA reagent. DharmaFECT achieved > 80% silencing at all siRNA concentrations, while HiPerFect (Qiagen) only achieved this level with PPIB at 100 nM. HeLa cells were transfected with SMARTpool TM reagents targeting Cyclophilin B (PPIB) or MAPKI at concentrations of 1, 5, 25, and 100 nM. mRNA expression (bars) was determined by branched DNA assay (Panomics Quantigene® Reagent System) and cell viability (data points) was determined by alamarBlue® (Biosource International).
Table adapted from Borawski et al. A study by scientists at Novartis concluded that in transfection optimization in 384-well format (in preparation for screening) one of the four DharmaFECT transfection reagents outperformed all other reagents in delivery efficiency and overall cell viability in 9 of 10 cell lines. Lipid transfection reagents tested were DharmaFECT 1-4, HiPerFect® (Qiagen), TransIT-TKO® (Mirus) Lipofectamine® 2000 and Oligofectamine® (Invitrogen). J. Borawski et al., Optimization Procedure for small interfering RNA Transfection in a 384-well format. J. Bimolecular Screening,12, 546-559 (2007).