Description:
PepMute™
siRNA Transfection Reagent, formulated from simulation of virus cell penetrating
peptides (CCPs), is a total novel
siRNA
delivery tool which provides more than 95% silencing efficiency at 1 nM siRNA in
variety of mammalian cells.
With our proprietary peptide simulation technology (PST),
hundreds of viral CCPs were simulated, synthesized and screened for gene delivery efficacy in variety
of mammalian cells (Figure 1). PepMute™
reagent was then identified and validated as an exceptionally efficient vector
for condensing and transfecting short (under 100 bp) single or double stranded
nucleic acids such as siRNA, miRNA mimics and DNA oligoes to wide spectrum of mammalian cells.
Figure 1. A cartoon showing PepMute™
siRNA Transfection Reagent was
developed by PST
Size:
-
PepMute™ Reagent,
1.0 ml, sufficient for ~1,333 reactions based on transfecting 0.5~5.0 pmol
siRNA or miRNA mimics in 24-well plate
- PepMute™ Transfection
Buffer, formulated for maximal transfection efficiency, 8.0 ml
Applications:
- siRNA miRNA mimics or mRNA transfection
- DNA/siRNA co-transfection
- DNA oligoes transfection
Storage:
Store at 4 °C and never freeze. If stored properly, the product is stable for 12 months or
longer.
Advantages:
- Excellent silencing at 1 nM siRNA
- Over 95% gene silencing in a wide variety of cells
- One tube reaction with easy standard protocol
- Compatible with serum and antibiotics
- Protocols adapted to HTS
- Very low cytotoxicity
- Very affordable
Silencing Efficacy Comparison between PepMute™ siRNA Transfection Reagent
and Leading Products
Figure 2.
Excellent silencing of endogenously expressed KIF11 (also known as EG5) in
HEK293 cells with 1.0 µl of PepMute™
reagent and 0.5 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. PepMute™ reagent effectively delivers EG5 siRNA (final 1.0 nM)
to HEK293 cells, leading to more than 80% of "round-up" phenotype of HEK293
cells 48h post transfection over negative control (final 1.0 nM with sham EG5 siRNA)
while leading siRNA transfectin reagents, Lipofectamine™ RNAiMAX
(RNAiMAX, 1.0 nM EG5 siRNA) / INTERFERin (1.0 nM EG5 siRNA) / Dharmafect (10.0 nM EG5 siRNA) and jetPRIME
(20 nM EG5 siRNA) give average 37%, 23%, 53% and 48% ball-shaped phenotype
respectively on HEK293 cells. The phenotype of "rounded-up" 293 cells
were visualized (upper panel) and quantified (lower panel) 48h post transfection with a Nikon
microscope.
Figure 3. Silencing efficiency comparison of PepMute™ Transfection Reagent
(upper panel) with Dharmafect 4 (middle panel) and Lipofectamine™
RNAiMAX (RNAiMAX, lower panel) siRNA
Transfection Reagents on A549 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 A549 cells growing on a 24-well plate.
Renilla luciferase activity was determined 36h 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
PepMute™ and Dharmafect™ 4 reagents delivered
significant gene silencing from 1.0 nM of renilla luciferase siRNA,
Lipofectamine™ RNAiMAX gave good knockdown only after
20 nM while enhanced gene
expression at low concentration of siRNA (0.5 and 1.0 nM respectively) was
observed.
Figure 4. PepMute™ Transfection Reagent knocked down stable GFP expression
in MCF7 cell (upper panel) and U2OS cell (lower panel) by reversely transfecting
5.0 and 1.0 nM GFP siRNA respectively.
Green fluorescence protein (GFP) was stably expressed in MCF7 and U2OS
cells. siRNA targeting
GFP gene (right panel) and a sham siRNA (left panel) were introduced into MCF7
and U2OS cells with final 5.0 and 1.0 nM respectively
by
reverse transfection with PepMute™ Transfection Reagent. GFP gene silencing was monitored
48h post transfection by
a Nikon fluorescence microscope. Quantitative analysis showed that
GFP siRNA at 5.0 and 1.0 nM delivered
by PepMute™ siRNA Transfection Reagent knocked down 90% and 95% stably expressed GFP in
MCF7 and U2OS cells respectively.
Data Sheet & Protocol
-
A Standard siRNA Transfection Protocol
-
A Reverse siRNA Transfection Protocol for HTS
-
A Standard siRNA/DNA
Co-transfection Protocol
