Thermo Scientific Accell eGFP Control siRNA is designed to target cloning vector pEGFP-N1 (Genbank accession # U55762) and has 100% homology with over 100 additional cloning vectors and transgenic eGFP sequences. Expressed green fluorescent protein (eGFP) is a reporter molecule for monitoring gene expression and protein localization in vivo and in real time. Accell eGFP control siRNA reagents silence expression of the GFP mRNA leading to a detectable decrease of fluorescence observed by fluorescent microscopy or by FACS analysis.
Highlights
• Accell siRNA enters cells without the need for transfection reagents, virus (or viral vectors), or instruments
• Novel siRNA modifications facilitate uptake, stability, specificity, and knockdown efficiency
• Target sequence: 5’ GCCACAACGTCTATATCAT 3’
Experimental considerations
• Accell siRNA works at a higher concentration than conventional siRNA; recommended 1 µM working concentration
• Delivery may be inhibited by the presence of BSA in serum. Optimization studies with serum-free media formulations (Accell Delivery Media) or < 2.5% serum in standard media is recommended
• Full-serum media can be added back after 48 hours of incubation. Optimal mRNA silencing is typically achieved by 72 hours or up to 96 hours for protein knockdown