Application

1) To evaluate the Simplicon™ expression system in targeted cell(s) before ordering a custom Simplicon™ RNA
2) To determine optimal transfection conditions to express the self-replicating RNA in cell lines

The Simplicon™ TagRFP RNA kit was developed to enable the evaluation of the Simplicon™ expression system in targeted cell(s) before ordering a custom Simplicon™ RNA. Simplicon™ TagRFP RNA kit may be used to determine optimal transfection conditions to express the self-replicating RNA in hard-to transfect somatic or primary cells.

Components

1. TagRFP RNA: (Part No. CS222753) One (1) vial containing 10 µL of RNA (1 ug/uL). Store at -80°C.

2. B18R RNA: (Part No. CS210584) One (1) vial containing 10 µL of RNA (1 ug/uL). Store at -80°C.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Simplicon™ is a novel system to effect immediate high sustained protein expression of the target gene(s) without the risk of integration. The technology employs a self-replicating RNA based on the Venezuelan equine encephalitis (VEE) genome. Because Simplicon™ is a synthetic RNA, there is no risk of the transgenes(s) integrating into the transfected cell′s genome. Moreover, the self-replicating nature of the Simplicon™ means that the target protein is continuously expressed in the presence of B18R protein and the selective agent, puromycin. The Simplicon™ technology has been successfully utilized for efficient human iPSC generation through the sustained expression of critical reprogramming factors, OKSG and OKSG-cMyc.
The Simplicon™ TagRFP RNA kit was developed to enable the evaluation of the Simplicon™ expression system in targeted cell(s) before ordering a custom Simplicon™ RNA. Simplicon™ TagRFP RNA kit may be used to determine optimal transfection conditions to express the self-replicating RNA in hard-to transfect somatic or primary cells. Simplicon™ TagRFP expresses a monomeric red (orange) fluorescent protein generated from the RFP of sea anemone Entacmaea quadricolor. TagRFP exhibits fluorescence withexcitation/emission maxima at 555/584 nm respectively, and brightness that is nearly three times higher than mCherry.

Legal Information

SIMPLICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to lot specific datasheet.

Quality

Discrete RNA band on RNA gel of appropriate size.
RFP-positive cells observed one day after transfection.
40-60% RFP-positive BJ human fibroblasts with FACS analysis.