GeneSilencer® Reagent Vs. siPORT NeoFX Reagent
HEK 293 Cells (click on images to enlarge)
Figure Legend: 293 HEK cells were transfected with 5, 10, 30 or 50 nM of Silencer® FAM Labeled
GAPDH siRNA with either GeneSilencer® (Genlantis, Inc.) or siPORT NeoFX (Ambion, Inc.) siRNA
transfection reagents according to the manufacturer's recommended protocols. Cells were incubated for
48 hours then fixed, permeablized, and incubated with AlexaFluor® 555 phalloidin (Invitrogen, Inc.), which
stains cellular actin red. Cells were then mounted on slides and stained with DAPI (Invitrogen, Inc.), which
stains the cell nuclei blue. Transfected cells were visualized by fluorescence microscopy using identical
exposure times for FITC, TRITC, and DAPI. The transfected GAPDH siRNA is localized in the cell cytosol
and can be seen as green fluorescent specks or dots.
HeLa S3 Cells (click on images to enlarge)
Figure Legend: HeLa S3 cells were transfected with 5, 10, 30 or 50 nM of Silencer® FAM Labeled GAPDH
siRNA with either GeneSilencer® (Genlantis, Inc.) or siPORT¿ NeoFX¿ (Ambion, Inc.) siRNA transfection
reagents according to the manufacturer's recommended protocols. Cells were incubated for 48 hours then
fixed, permeablized, and incubated with AlexaFluor® 555 phalloidin (Invitrogen, Inc.), which stains cellular
actin red. Cells were then mounted on slides and stained with DAPI (Invitrogen, Inc.), which stains the cell
nuclei blue. Transfected cells were visualized by fluorescence microscopy using identical exposure times for
FITC, TRITC, and DAPI. The transfected GAPDH siRNA is localized in the cell cytosol and can be seen as
green fluorescent specks or dots.
Description
GeneSilencer® siRNA Transfection Reagent is a novel cationic lipid formulation specifically designed for efficient delivery of siRNAs (small interfering RNAs) into a wide variety of cell types. siRNAs are short, gene-specific double-stranded RNAs that can cause gene silencing in mammalian cells by catalytically cleaving greater than 95% of the target mRNA (1,2,3).
Contents
- Hydrated GeneSilencer® Lipid (1 vial x 750 µl for T500750 or 5 vials x 750 µl for T505750)
- siRNA Diluent (1 tube x 4 ml for T500750 / 5 tubes x 4 ml for T505750)
Storage
The GeneSilencer® siRNA Transfection Reagent is shipped at room temperature. For maximum stability, store all reagents at 4ºC upon receipt. If stored properly, all components are stable for 6 months.
References
- Elbashir, S.M. et al. (2001) Nature 411: 494-498.
- Caplen, N.J. et al. (2001) Proc Natl Acad Sci USA 98: 9742-9747.
- Sharp, P.A. (2001) Genes and Development 15: 485-490.
" In contrast [to the Mouse Nucleofector kit from Amaxa Biosystems and transfection reagents from Santa Cruz Biotechnology], the most effective method was the lipid-based method using the siRNA transfection reagent GeneSilencer from Genlantis. This protocol resulted in up to 92% FITC-siRNA positive cells after 4 h which declined to 62% and 59% 24 and 48 h post-transfection, respectively. The transfected BM-DC remained CD11c positive, expressed high MHC class II and intermediate CD40 and were functional as APC.."
Pedersen, CD, Fang, JJ, and Pedersen, AE, (2009) A comparative study of transfection methods for RNA interference in bone marrow-derived murine dendritic cells. Scand J Immunol 70(5): 447.
GeneSilencer Primary Cell Citations
