GenMute Reagent is a novel biodegradable non-liposomal siRNA/miRNA & DNA delivery tool. With our proprietary pH Dependent Conformational Change (PDCC) technology, the biodegradable transfection reagent was formulated by addition of pre-screened hydrophobic groups to its side chain, making GenMute Reagent a versatile and most powerful gene delivery tool in the market. GenMute Reagent have been validated to effectively and reproducibly transfect single siRNA/miRNA or co-transfect DNA/siRNA or DNA/miRNA to variety of mammalian cells.
Advantages:
- Excellent silencing at low concentrations with only 1.0 nM siRNA / miRNA mimics
- Minimizing non-specific and off target effects in the cell
- One tube reaction
- Best for broad mammalian cells
- Very low cytotoxicity
- Very affordable
Comparisons of Silencing Efficacy of GenMute siRNA & DNA Transfection Reagent with Brand Name Products
Knockdown efficacy comparison of GenMute Transfection Reagent (upper panel) vs. Dharmafect 4 siRNA Transfection Reagent (middle panel) and Lipofectamine 2000 (lower panel) on HEK293 cells. siRNA targeting renilla luciferase at different final concentrations ranging from 0.5 to 20 nM was co-transfected with renilla luciferase gene (0.5 µg of pRL-CMV DNA per well) by the above three transfection reagents per manufacturers' protocols into HEK293 cells growing on a 24-well plate. Renilla luciferase activity was determined 24h after post co-transfection with renilla luciferase determination system (Promega). The luminescence was measured from 5.0 µl of lysate during 10s integration with a luminometer (Beckman Coulter LD 400). Luciferase activity was expressed as light units integrated over 10s (RLU) and normalized per mg of cell protein by using the BCA assay. The errors bars represent standard deviation derived from triplicate experiments. Luciferase-silencing efficiency was calculated relative to untreated cells. While GenMute and Dharmafect 4 reagents delivered significant gene silencing from 1.0 nM of renilla luciferase siRNA, lipofectamine 2000 gave good knockdown only after 30 nM (data not shown).
Excellent silencing of endogenously expressed KIF11 (also known as Eg5) in HEK293 cells with 0.5 µl of GenMute reagent and 5.0 pmol Eg5 siRNA per well of 24-well plate. KIF11 (also known as Eg5) encodes a motor protein that belongs to the kinesin-like protein family involved in chromosome positioning and bipolar spindle formation during cell mitosis. A reduction in KIF11 levels causes mitotic arrest. GenMute reagent effectively delivers Eg5 siRNA (final 10 nM, right panel) to HEK293 cells at only 0.5 µl per well of 24-well plate vs. negative control (final 10 nM. left panel). Compared with negative control (left panel), phenotype of "rounded-up" 293 cells were visualized 24 hours post transfection (right panel) with a Nikon microscope.
GenMute Transfection Reagent knocked down endogenous lamin A/C gene expression in Hela cells. A siRNA targeting lamin A/C gene (right panel) and a sham siRNA (left panel) were introduced into Hela cells (final 1.0 nM) by GenMute Transfection Reagent. Lamin A/C gene silencing was monitored 24h post transfection by immunofluorescence. Lamin A/C was probed with a mouse monoclonal lamin A/C specific antibody followed by addition of FITC-conjugated anti-mouse antibodies. Quantitative analysis showed that lamin siRNA at 1.0 nM delivered by GenMute Transfection Reagent knocked down 94% endogenously expressed lamin A/C in Hela cells.
