GenMute™ siRNA试剂是一种新型的可生物降解的非脂质体的siRNA / miRNA和DNA的载体工具。运用我们专有的PH依赖构象变化技术，这款可生物降解的转染试剂在它的侧链上添加了预筛选出来的疏水集团，使得GenMute™试剂成为市场上最万能的最强大的基因载体工具。GenMute™试剂，已被证实能有效的，可重复性转染单siRNA片段和miRNA模拟物或共转染DNA/siRNA片段或DNA/miRNA片段到各种哺乳动物细胞中。
- GenMute™转染试剂，1.0ml，足够在24孔板中转染5.0 pmoles的siRNA或miRNA模拟物达到2000个反应。
- GenMute™ 转染试剂缓冲液（5X），与PGenMute™ 试剂配合使用以获得转染效率最大化，8.0ml（5X）浓缩液就可以制成40ml的工作液体。
- 对各种哺乳动物细胞的单siRNA 转染或者 DNA/siRNA 共转染。
- 对各种哺乳动物细胞的单 miRNA， miRNA模拟物 或者 miRNA抑制剂的转染或者 DNA/miRNA 或DNA/miRNA模拟物的共转染。
- 在只有 1.0 nM siRNA / miRNA模拟物的低浓度中，也能达到非常好的沉默效应。
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 thatlamin siRNA at 1.0 nM delivered by GenMute™ Transfection Reagent knocked down 94% endogenously expressed lamin A/C in Hela cells.