mRNA Reprogramming Factors Set: hOKSML
Stemgent mRNA Reprogramming Factors Set: hOKSML contains mRNA encoding Oct4, Klf4, Sox2, c-Myc, Lin-28, and nuclear GFP (nGFP). These in vitro transcribed mRNAs incorporate both pseudouridine and 5-methylcytidine-modified nucleotides to minimize the cellular interferon response to single stranded RNA.1,2 Repeated transfection of mRNA encoding Oct4, Klf4, Sox2, c-Myc and Lin-28 has been shown to reprogram human fibroblasts.2 nGFP mRNA is included for use as a positive control for RNA transfection during the cellular reprogramming process.
- Faster reprogramming, colonies emerge as early as 16 days
- Stoichiometric control of individual reprogramming factors
- Eliminates need for backend screening of viral systems
- Validated protocol using normal and diseased cells
Kit / Set Components
2 vials - Stemgent Oct4 mRNA, Human (Cat. No. 05-0014), 20 μg
1 vial - Stemgent Klf4 mRNA, Human (Cat. No. 05-0015), 20 μg
1 vial - Stemgent Sox2, Human (Cat. No. 05-0016), 20 μg
1 vial - Stemgent c-Myc mRNA, Human (Cat. No. 05-0018), 20 μg
1 vial - Stemgent Lin-28 mRNA, Human (Cat. No. 05-0017), 20 μg
1 vial - Stemgent nGFP mRNA (Cat. No. 05-0019), 20 μg
100 ng/μl in RNase-free TE buffer, pH 7.0
Storage and Stability
Store at or below -70°C. Stable for a minimum of 3 months from date of receipt when stored as directed. Wear gloves when handling this product. Use RNase-free tubes and barrier pipette tips. Thaw on ice. Keep on ice until the sample is removed. Avoid multiple freezing and thawing cycles (<5). Aliquot to experimental quantities.
Proper protein expression and localization have been confirmed by immunocytochemistry for reprogramming factors and by fluorescence microscopy for nGFP.
mRNAs should be transfected using a protocol specifically optimized for the cell type used in the experiment.
- Stemgent® Oct4 mRNA, human (Cat.No.05-0014 )
- Stemgent® Klf4 mRNA, Human (Cat.No.05-0015)
- Stemgent® Sox2 mRNA, Human (Cat.No.05-0016)
- Stemgent® c-Myc mRNA, Human (Cat.No.05-0018)
- Stemgent® Lin-28 mRNA, Human (Cat.No.05-0017)
- Stemgent® nGFP mRNA (Cat.No.05-0019)
- Pluriton™ mRNA Reprogramming Medium (Cat.No. 00-0070)
- Stemfect™ RNA Transfection Kit (Cat.No.00-0069)
- Stemolecule™ Y27632 (Cat.No.04-0012)
- Stemfactor™ Fibroblast Growth Factor-basic (Human Recombinant) (Cat.No.03-0002)
Safety Data Sheets
- Generation of Safe, Integration-free Human Induced Pluripotent Stem (iPS) Cells using the mRNA Reprogramming System
- Scientific Poster: mRNA Reprogramming SLAS 2011
- FAQs: mRNA Reprogramming
- Limited Use License MIT
- Limited Use License iPS Academia Japan
- Angel, M., and Yanik, M.F. (2010) Innate immune suppression enables frequent transfection with RNA encoding reprogramming proteins. PLoS One 5: e11756.
- Warren, L., Manos, P.D., Ahfeldt, T., Loh, Y.H., Li, H., Lau, F., Ebina, W., Mandal, P.K., Smith, Z.D., Meissner, A., Daley, D.Q., Brack, A.S., Collins, J.J., Cowan, C., Schlaeger, T.M., and Rossi, D.J. (2010) Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic modified mRNA. Cell Stem Cell 7: 618-630.
- Yakubov, E., Rechavi, G., Rozenblatt, S., and Givol, D. (2010) Reprogramming of human fibroblasts to pluripotent stem cells using mRNA of four transcription factors. Biochem Biophys Res Commun. 394: 189-193.
- Pankaj K Mandal and Derrick J Rossi. (2013) Reprogramming human fibroblasts to pluripotency using modified mRNA. Nature Protocols, 8: 568 – 582.
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