SCR550 Simplicon™ RNA Reprogramming Kit (OKSG)

SCR550
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      Overview

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      Acheive 5-fold increase in IPS cell generation using NEW RiboJuice mRNA Transfection Reagent!
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      Description
      Catalogue NumberSCR550
      DescriptionSimplicon™ RNA Reprogramming Kit (OKSG)
      Background InformationVarious methods utilizing viruses, DNA, RNA, miRNA and protein have been developed to generate integration-free induced pluripotent stem cells (iPSCs). Disadvantages to existing methods include: (1) low reprogramming efficiency (i.e. DNA and protein), (2) a lengthy requirement for negative selection and subcloing steps to remove persistent traces of the virus (i.e. Sendai virus)1 and (3) for daily transfections of four individual in vitro generated mRNAs over a 14 day period (i.e. mRNA based)2.

      EMD Millipore’s Simplicon™ RNA Reprogramming Kit is a safe and efficient method to generate integration free, virus-free human iPS cell using a single transfection step. The technology is based upon a positive strand, single-stranded RNA species derived from non-infectious (non-packaging), self-replicating Venezuelian equine encephalitis (VEE) virus3. The Simplicon RNA replicon is a synthetic in vitro transcribed RNA expressing all four reprogramming factors (OKS-iG; Oct4, Klf4, Sox2 and Glis1) in a polycistronic transcript that is able to self-replicate for a limited number of cell divisions.

      View our Application Note! (See the Data!)
      References
      Product Information
      Components
      • 1. VEE-OKS-iG RNA: (Part No. CS210583) 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.
      • 3. Human Recombinant B18R Protein, Carrier-Free: (Part No. GF156) One (1) vial containing 50 ug of 0.5 mg/mL stock of B18R protein. Store at -80°C.
      • 4. Human iPS Reprogramming Boost Supplement II: (Part No. SCM094). Store at -20°C.
      Applications
      ApplicationEMD Millipore’s Simplicon™ RNA Reprogramming Kit is a safe and efficient method to generate integration free, virus-free human iPS cell using a single transfection step of self-replicating synthetic RNA for Oct4, Klf4, Sox2 and Glis1.
      Application NotesAdvantages of the Simplicon™ RNA Reprogramming Kit:

      • Integration-free, footprint-free iPS cells. No risk of genomic integration
      • Safe, virus free, synthetic polycistronic RNA replicon (all four reprogramming factors in 1 RNA strand)
      •Only 1-day transfection required. The RNA replicon is able to self-replicate, elliminating the need for laborious daily transfection of multiple individual mRNAs over a 14 day period.
      • Efficient and rapid reprogramming.
      • No screening required to ensure viral remnants are not present.
      • Controlled elimination of synthetic RNA replicon by removal of B18R protein.
      • Validated for reprogramming in feeder-based and feeder-free culture conditions.
      Biological Information
      Stem Cell Type
      • Induced Pluripotent Stem Cells
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceTested to confirm the generation of iPS cells from p6 human foreskin fibroblasts. Other cell types have not been tested and thus similar results can not be guaranteed.
      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 Conditions• VEE-OKS-iG and B18R RNAs: Stable for 4 months from date of receipt when stored appropriately at -80°C. For best recovery, quick-spin the vial prior to opening. Thaw on ice. While on ice, aliquot into sterile, nuclease-free eppendorf tubes and store at -80°C. Limit repeated freeze-thaw cycles. Use in a sterile RNAse-free environment.

      • Human recombinant B18R protein: Stable for 4 months from date of receipt when stored appropriately at -80°C. For best recovery, quick-spin the vial prior to opening. Thaw on ice. While on ice, aliquot B18R protein into sterile, nuclease-free, low protein-binding eppendorf tubes and store at -80°C. B18R protein must be kept on ice in order to avoid degradation. Limit repeated freeze-thaw cycles. Use in a sterile RNAse-free environment.

      • Human iPS Reprogramming Boost Supplement II: Stable for 4 months at -20˚C from date of receipt. Upon first thaw, aliquot into smaller working volumes and freeze at -20°C. Upon addition of the small molecule components to the media, filter the supplemented media with a 0.22 μM filtration unit and stored at 2-8˚C. For optimal results, prepare sufficient supplemented media for a 1 week supply of media changes.
      Packaging Information
      Material Size1 kit
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Simplicon™ RNA Reprogramming Kit (OKSG) SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Renal Subcapsular Transplantation of PSC-Derived Kidney Organoids Induces Neo-vasculogenesis and Significant Glomerular and Tubular Maturation In Vivo.
      van den Berg, CW; Ritsma, L; Avramut, MC; Wiersma, LE; van den Berg, BM; Leuning, DG; Lievers, E; Koning, M; Vanslambrouck, JM; Koster, AJ; Howden, SE; Takasato, M; Little, MH; Rabelink, TJ
      Stem Cell Reports  10  751-765  2018

      Show Abstract
      29503086 29503086
      Characterization of induced tissue-specific stem cells from pancreas by a synthetic self-replicative RNA.
      Miyagi-Shiohira, C; Nakashima, Y; Kobayashi, N; Saitoh, I; Watanabe, M; Noguchi, H
      Sci Rep  8  12341  2018

      Show Abstract
      30120295 30120295
      An integration-free, virus-free rhesus macaque induced pluripotent stem cell line (riPSC90) from embryonic fibroblasts.
      Sosa, E; Kim, R; Rojas, EJ; Hosohama, L; Hennebold, JD; Orwig, KE; Clark, AT
      Stem Cell Res  21  5-8  2017

      Show Abstract
      28677539 28677539

      Brochure

      Title
      Hallmarks of Aging (EMD)
      Simplifying the iPSC Workflow

      Technical Info

      Title
      Single transfection of a synthetic polycistronic self-replicative RNA yields high numbers of transgene-free human iPSCs

      User Guides

      Title
      SimpliconTM RNA Reprogramming Kit