DharmaFECT Set of 4 Transfection Reagents includes one vial of each chemically distinct DharmaFECT formulation, and is offered to help determine the appropriate transfection reagent for any cell types where a recommended formulation is not available, or when experimental conditions call for re-optimization of siRNA or microRNA transfection. Our online resources for over 35 cell lines can help determine the best single DharmaFECT Transfection Reagent formulation for your research, but if you are unable to find recommendations for your cell line, it is advisable to optimize delivery with DharmaFECT 1, 2, 3, and 4 Transfection Reagents.
- Four distinct formulations for transfection optimization studies in new cell types or experimental conditions
- Optimized for small RNA (siRNA or microRNA) delivery at low concentrations
- Low toxicity – Maintain cell viability with formulations specially formulated to deliver siRNA
While DharmaFECT 1 is the most all-purpose transfection reagent (demonstrating efficient, low-toxicity delivery to over 80% of validated cell types), DharmaFECT 2, 3, and 4 are available to support a wider range of cell types. Distinct formulations permit more thorough optimization of transfection for novel cell types, high-value experiments and RNAi screening.
Consult the DharmaFECT Cell Type Guide of transfection optimization results for the DharmaFECT formulation that is recommended for your cell type.
Editing of PSMD7 gene in U2OS-(Ubi)EGFP cells using Edit-R Cas9 Nuclease protein NLS delivered by DharmaFECT transfection reagents
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).
Editing of VEGFA gene in HEK293T cells using Edit-R Cas9 Nuclease mRNA delivered by DharmaFECT transfection reagents
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 provide an improved dynamic range
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 (100 nM) using a range of volumes (0.05 to 1.6 μL/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).
DharmaFECT Transfection Reagents provide highly efficient delivery at low siRNA concentrations
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).
DharmaFECT outperforms other reagents in optimization study
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).