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SCR721 TagRFP Simplicon® RNA (E3L) Kit

To determine optimal transfection conditions to express the self-replicating RNA of your interest through using Simplicon® Cloning Vector (E3L) (Part #:SCR724) in hard-to- transfect somatic or primary cells

TagRFP Simplicon® RNA (E3L) Kit MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
SCR721
1 kit  
Purchase on Sigma-Aldrich

Áttekintés

Replacement Information
Description
Catalogue NumberSCR721
DescriptionTagRFP Simplicon® RNA (E3L) Kit
Alternate Names
  • Self-replicating TagRFP RNA
Background InformationSimplicon® is a novel system to effect immediate high sustained protein expression of multiple genes into transfected cells without the risk of genome integration. The technology employs a single, synthetic, polycistronic, self-replicating RNA based on the Venezuelan equine encephalitis (VEE) genome1,2,3. The Simplicon® RNA contains only genes encoding the VEE RNA replication machinery while the structural proteins that are required to make an infectious particle have been removed and replaced with the transgenes of interest.

Introduction and replication of the Simplicon® RNA is expected to elicit a strong interferon response in transfected cells. To suppress the IFN responses, a Vaccinia virus protein4, B18R, is used for the original Simplicon® technology. Recently, we found that another Vaccinia virus protein4, E3L, also suppresses the IFN responses in Simplicon® RNA expression. B18R neutralizes type I interferons by direct binding, while E3L inhibits the cytoplasmic signaling pathwaycs of IFN responses. Therefore, B18R and E3L are both employed in the Simplicon® Expression System and work collaboratively to suppress IFN responses. As a result, there is increased cell viability during RNA transfection and increased expression of the transgenes. The Simplicon® Expression System works in human cells and is not expected to work in mouse cells. This is because the B18R does not effectively neutralize mouse interferon (IFN)-β.

The TagRFP Simplicon® RNA (E3L) Kit was developed to enable the evaluation of the Simplicon® Expression System in targeted cells. E3L4 incorporation in the Simplicon® Expression System strongly suppresses interferon responses and enables increased expression of the target gene(s). TagRFP Simplicon® RNA expresses a monomeric red (orange) fluorescent protein generated from the RFP of sea anemone Entacmaea quadricolor. TagRFP exhibits fluorescence with excitation/ emission maxima at 555/584 nm respectively, and brightness that is nearly three times higher than mCherry5.6.
References
Product Information
Components
  • 1.TagRFP Simplicon® RNA (E3L): (Cat. # CS224550) One (1) vial containing 10 µL of RNA (1 µg/µL). Store at -80°C.

    2.B18R-E3L RNA: (Cat. # CS224503) One (1) vial containing 10 µL of RNA (1 µg/µL). Store at -80°C.
Applications
ApplicationTo determine optimal transfection conditions to express the self-replicating RNA of your interest through using Simplicon® Cloning Vector (E3L) (Part #:SCR724) in hard-to- transfect somatic or primary cells
Application NotesThe Simplicon® TagRFP RNA (E3L) Kit was developed to enable the evaluation of the Simplicon® expression system in your cell line(s). For example, it may be used to determine optimal transfection conditions to express the self-replicating RNA of your interest through using Simplicon® Cloning Vector (E3L) (Part #:SCR724) in hard-to- transfect somatic or primary cells.
Biological Information
ConcentrationPlease refer to lot specific datasheet.
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Quality Assurance•Discrete RNA band on RNA gel of appropriate size.
•40-60% RFP-positive human foreskin fibroblasts (HFFs) with FACS analysis.
Usage Statement
  • 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.
Storage and Shipping Information
Storage Conditions1.TagRFP Simplicon® RNA (E3L): (Cat. # CS224550) One (1) vial containing 10 µL of RNA (1 µg/µL). Store at -80°C.

2.B18R-E3L RNA: (Cat. #CS224503) One (1) vial containing 10 µL of RNA (1 µg/µL). Store at -80°C.
Packaging Information
Material Size1 kit
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Katalógusszám GTIN
SCR721 04054839501715