PepMute™ Plus siRNA Transfection Reagent

Description:
PepMute™ Plus siRNA & DNA Transfection Reagent is an upgraded version of PepMute™ Reagent (Cat # SL100566), enhanced by crosslinking pre-screened hydrophobic amino acids to its peptide backbone. Leveraging our proprietary pH-Dependent Conformational Change (PDCC) technology, the 36–amino-acid peptide backbone has been modified with hydrophobic amino acids at all amine groups. This design allows PepMute™ Plus to self-assemble during transfection complex formation, resulting in a highly versatile and powerful gene delivery platform.

PepMute™ Plus provides over 95% silencing efficiency at 1 nM siRNA across a wide range of mammalian cell types. The reagent has been validated for efficient and reproducible transfection of single DNA, single siRNA, or co-transfection of DNA and siRNA in various mammalian cells.

Figure 1. A cartoon showing PepMute™ Plus siRNA Transfection Reagent was developed

Size & content:
– PepMute™ Plus Transfection Reagent, 1.0 mL, sufficient for ~1000 reactions based on transfecting 2.5 pmols siRNA in 24-well plate
– PepMute™ Plus Transfection Buffer(5x ), formulated for maximal transfection efficiency, 8.0 mL

Storage:
Store at 4 °C for PepMute™ Plus Reagent and RT for PepMute™ Transfection Buffer (5x ). If stored properly, the product is stable for 12 months or longer.

Advantages:
– Effective knockdown (95%) with only 0.5 pmol siRNA (1.0 nM siRNA in 24-well plate)
– Formulated for hard-to-transfect mammalian cells
– One tube reaction and easy transfection protocol
– Best for broad spectrum, especially hard-to-transfect mammalian cells
– Very low cytotoxicity

Comparisons of Knockdown Efficacy of PepMute™ Plus siRNA & DNA Transfection Reagent with Brand Name Products

Figure 2. Knockdown efficacy comparison of PepMute™ Plus 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 the renilla luciferase gene (0.5 µg of pRL-CMV DNA per well) by the above three transfection reagents per manufacturer’s protocols into HEK293 cells growing on a 24-well plate. Renilla luciferase activity was determined 24h after post-transfection with the 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 error bars represent the standard deviation derived from triplicate experiments. Luciferase-silencing efficiency was calculated relative to untreated cells. While PepMute™ Plus 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).

Figure 3. PepMute™ Plus Transfection Reagent knocked down GFP gene expression in HepG2 cells. GFP cDNA, pEGFP-N3, was co-transfected with a siRNA targeting the GFP gene (final 5.0 nM, right panel) and a sham siRNA (final 5.0 nM, left panel) into HepG2 cells by PepMute™ Plus Transfection Reagent. GFP fluorescence was visualized 24h post-transfection with a Nikon T2000 fluorescence microscope. Quantitative analysis showed that GFP siRNA at 5.0 nM delivered by PepMute™ Plus Transfection Reagent knocked down 96% co-transfected GFP expression in HepG2 cells.

Data Sheet & Protocol
– A Protocol for siRNA Transfection and DNA/siRNA Co-Transfection
– A Short Protocol for siRNA Transfection for Advanced Users 
– A Short Protocol for siRNA/DNA Co-transfection for Advanced Users 
– A Protocol for DNA Transfection to Mammalian Cells